Purinone derivative

ABSTRACT

Compounds represented by general formula (I) (all of the symbols in the formula conform to the definitions in the Description) are compounds that, in addition to having a Btk-selective inhibitory activity, exhibit an excellent metabolic stability and can avoid hepatotoxicity or the like, and as a consequence can provide safe therapeutic agents for diseases in which B cells or mast cells participate.

TECHNICAL FIELD

The present invention relates to compounds represented by generalformula (I)

(all of the symbols in the formula have the same definitions as givenbelow), optical isomers thereof or their mixture, salts thereof,solvates thereof, N-oxides thereof, and prodrugs thereof (abbreviatedbelow as “compounds of the present invention”).

BACKGROUND ART

Bruton's tyrosine kinase (abbreviated below as “Btk”) belongs to the Tecfamily of kinases, which are non-receptor tyrosine kinases, and isselectively expressed in the B cell and myelocyte lines. Btk plays animportant role in signal transduction in B cells and is a factor thatcontributes to the survival, differentiation, proliferation, andactivation of B cells. Signaling in B cells via the B cell antigenreceptor (BCR) induces a broad range of biological responses, andabnormal signal transduction here causes abnormal B cell activation andthe formation of pathogenic autoantibodies. Btk is believed to form alink in the BCR-mediated signal transduction pathways into B cells.Thus, X-linked agammaglobulinemia (XLA) is known to be caused by adefect in the human Btk gene that results in the induction of abnormal Bcell differentiation and a drastic decline in immunoglobulin production(refer to Non-patent Document 1). The symptoms of this disease include asubstantial decline in B cells in the peripheral blood and an increasedsusceptibility to bacterial infections. Btk is also known to participatein mast cell activation and in the physiological functions of platelets.Due to this, compounds that have a Btk inhibitory activity are effectivefor the treatment of diseases in which B cells or mast cellsparticipate, for example, allergic diseases, autoimmune diseases,inflammatory diseases, thromboembolic diseases, and cancers (refer toNon-patent Document 2).

The following compounds are known as prior art for the compounds of thepresent invention.

Compounds represented by general formula (A) are known as compounds thathave a Btk inhibitory activity

(in the formula, L_(a) ^(A) represents CH₂, O, NH, or S; Ar^(A)represents substituted or unsubstituted aryl or substituted orunsubstituted heteroaryl; Y^(A) represents any substituent selected fromalkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl;Z^(A) represents CO, OCO, NHCO, or CS; R^(7-A) and R^(8-A) eachindependently represent H, unsubstituted C₁-C₄ alkyl, substituted C₁-C₄alkyl, unsubstituted C₁-C₄ heteroalkyl, substituted C₁-C₄ heteroalkyl,unsubstituted C₃-C₆ cycloalkyl, substituted C₃-C₆ cycloalkyl,unsubstituted C₂-C₆ heterocycloalkyl, and substituted C₂-C₆heterocycloalkyl; or R^(7-A) and R^(8-A) together form a bond; andR^(6-A) represents H, substituted or unsubstituted C₁-C₄ alkyl,substituted or unsubstituted C₁-C₄ heteroalkyl, C₁-C₆ alkoxyalkyl, C₁-C₈alkylaminoalkyl, substituted or unsubstituted C₃-C₆ cycloalkyl, orsubstituted or unsubstituted aryl (the definitions of these groups havebeen excerpted))(refer to Patent Documents 1, 2, and 3).

On the other hand, for example, compounds represented by general formula(B)

(in the formula, Q^(1B) and Q^(2B) are independently selected fromCX^(1B), CX^(2B), and nitrogen; Q^(3B) represents N or CH; X^(1B) andX^(2B) are independently selected from the group consisting of hydrogen,(C₁-C₆) alkyl, cyano, halogen, and so forth; R^(1B) is selected from thegroup consisting of hydrogen and (C₁-C₆) alkyl; yB represents 0 or aninteger from 1 to 3; R^(2B) and R^(3B) are independently selected fromhydrogen and (C₁-C₆) alkyl; R^(4B) is selected from the group consistingof alkyl, heterocyclyl, aryl, heteroaryl, and so forth; and R^(5B) isselected from the group consisting of alkyl, heterocyclyl, andsubstituted heterocyclyl (the definitions of these groups have beenexcerpted)) (refer to Patent Document 4) are known as compounds thathave a purinone skeleton.

Compounds represented by general formula (C) are also known

(in the formula, X^(C) is selected from the group consisting of nitrogenand CR^(8C); R^(8C) is selected from the group consisting of hydrogen,halogen, substituted or unsubstituted alkyl, and so forth; Q^(1C) isselected from the group consisting of O, S, and so forth; Z^(C) isselected from the group consisting of oxygen, sulfur, and NY^(5C);Y^(5C) is selected from the group consisting of hydrogen, substituted orunsubstituted alkyl, and so forth; Q^(2C), Q^(3C), and Q^(4C) areindependently selected from the group consisting of hydrogen,substituted or unsubstituted alkyl, substituted or unsubstituted aryl,and so forth; R^(2C) is selected from the group consisting of hydrogenand substituted or unsubstituted alkyl; and nC represents 0, 1, 2, 3, or4 (the definitions of these groups have been excerpted)) (refer toPatent Document 5).

Compounds having a purinone skeleton are also disclosed as formula 20(refer to paragraph number 0028) in Patent Document 6.

Compounds represented by general formula (D) are also known

(in the formula, R^(1D) represents a group selected from hydrogen,substituted or unsubstituted alkyl,

and so forth; R^(2D) represents substituted or unsubstituted alkyl,substituted or unsubstituted alkenyl, or substituted or unsubstitutedalkynyl; Y^(D) represents a group selected from O, C—NO₂, and S; Z^(D)represents a group selected from

and so forth; here, A^(D), D^(D), E^(D), and M^(D) each independentlyrepresent CR^(12D), N, and N-oxide; R^(12D) represents a group selectedfrom hydrogen, halogen, amino, hydroxy, and cyano; X^(D) represents agroup selected from O, C—NO₂, and S; R^(6D) represents a group selectedfrom substituted or unsubstituted cycloalkyl, substituted orunsubstituted heteroalkyl, and substituted or unsubstituted aryl; theindication aD represented by the dashed line represents a single bond ora double bond; and nD represents an integer selected from 0, 1, and 2)(refer to Patent Document 7).

The compounds of the present invention are compounds that, in additionto having a Btk-selective inhibitory activity, exhibit an excellentmetabolic stability and can avoid a CYP inhibitory activity and adversereactions such as, for example, hepatotoxicity; however, there is nostatement or suggestion relating to these characteristic features in anyof the prior art documents.

-   Patent Document 1: Japanese Translation of PCT Application No.    2010-504324-   Patent Document 2: WO 2008/121742-   Patent Document 3: WO 2010/009342-   Patent Document 4: WO 2008/060301-   Patent Document 5: WO 2007/142755-   Patent Document 6: Japanese Translation of PCT Application No.    2003-509427-   Patent Document 7: WO 2003/037890-   Non-patent Document 1: Nature, Volume 361, pp. 226-233, 1993-   Non-patent Document 2: Anticancer Agents in Medicinal Chemistry,    Volume 7, Number 6, pp. 624-632, 2007

DISCLOSURE OF THE INVENTION

In order to provide a very safe therapeutic agent for diseases in whichB cells and/or mast cells participate, an object of the presentinvention is to develop a compound that, in addition to having aBtk-selective inhibitory activity, exhibits an excellent metabolicstability and can avoid hepatotoxicity or the like.

In order to achieve the above object, the present inventors carried outintensive investigations directed to discover compounds having aBtk-selective inhibitory activity and discovered the compounds of thepresent invention as a result. Moreover, it was also discovered thatthese compounds are compounds that exhibit an excellent metabolicactivity and can avoid hepatotoxicity or the like, and the presentinvention was achieved as a result.

That is, the present invention relates to

[1] a compound represented by general formula (I)

(in the formula,L represents (1) —O—, (2) —S—, (3) —SO—, (4) —SO₂—(5) —NH—, (6) —C(O)—,(7) —CH₂—O—, (8) —O—CH₂—, (9) —CH₂—, or (10) —CH(OH)—;R¹ represents (1) a halogen atom, (2) a C₁₋₄ alkyl group, (3) a C₁₋₄alkoxy group, (4) a C₁₋₄ haloalkyl group, or (5) a C₁₋₄ haloalkoxygroup;ring1 represents a 4- to 7-membered cyclic group, which may besubstituted by from one to five substituents each independently selectedfrom the group consisting of (1) halogen atoms, (2) C₁₋₄ alkyl groups,(3) C₁₋₄ alkoxy groups, (4) nitrile, (5) C₁₋₄ haloalkyl groups, and (6)C₁₋₄ haloalkoxy groups, wherein when two or more substituents arepresent on ring1, these substituents may form a 4- to 7-membered cyclicgroup together with the atoms in ring1 to which these substituents arebound;ring2 represents a 4- to 7-membered saturated heterocycle, which may besubstituted by from one to three —K—R²;K represents (1) a bond, (2) a C₁₋₄ alkylene, (3) —C(O)—, (4)—C(O)—CH₂—, (5) —CH₂—C(O)—, (6) —C(O)O—, or (7) —SO₂— (wherein the bondon the left is bound to the ring2);R² represents (1) a C₁₋₄ alkyl, (2) a C₂₋₄ alkenyl, or (3) a C₂₋₄alkynyl group, each of which may be substituted by from one to fivesubstituents each independently selected from the group consisting of(1) NR³R⁴, (2) halogen atoms, (3) CONR⁵R⁶, (4) CO₂R⁷, and (5) OR⁸;R³ and R⁴ each independently represent (1) a hydrogen atom, or (2) aC₁₋₄ alkyl group which may be substituted by OR⁹ or CONR¹⁰R¹¹;R³ and R⁴ may, together with the nitrogen atom to which they are bound,form a 4- to 7-membered nitrogenous saturated heterocycle, which may besubstituted by an oxo group or a hydroxyl group;R⁵ and R⁶ each independently represent (1) a hydrogen atom, (2) a C₁₋₄alkyl group, or (3) a phenyl group;R⁷ represents (1) a hydrogen atom or (2) a C₁₋₄ alkyl group;R⁸ represents (1) a hydrogen atom, (2) a C₁₋₄ alkyl group, (3) a phenylgroup, or (4) a benzotriazolyl group;R⁹ represents (1) a hydrogen atom or (2) a C₁₋₄ alkyl group;R¹⁰ and R¹¹ each independently represent (1) a hydrogen atom or (2) aC₁₋₄ alkyl group;n represents an integer from 0 to 4;m represents an integer from 0 to 2; andwhen n is two or more, the R¹'s may be the same as each other or maydiffer from one another),an optical isomer thereof or their mixture, a salt thereof, a solvatethereof, an N-oxide thereof, or a prodrug thereof;

[2] the compound according to [1] above, wherein R² is a C₂₋₄ alkenylgroup or a C₂₋₄ alkynyl group, each of which may be substituted by fromone to five substituents each independently selected from the groupconsisting of (1) NR³R⁴, (2) halogen atoms, (3) CONR⁵R⁶, (4) CO₂R⁷, and(5) OR⁸;

[3] the compound according to [1] above, wherein ring1 is a benzene,cyclohexane, or pyridine ring, each of which may be substituted by fromone to five substituents each independently selected from the groupconsisting of (1) halogen atoms, (2) C₁₋₄ alkyl groups, (3) C₁₋₄ alkoxygroups, (4) nitrile, and (5) CF₃;

[4] the compound according to [1] above, wherein ring2 is a 4- to7-membered nitrogenous saturated heterocycle, which may be substitutedby from one to three —K—R²;

[5] the compound according to [4] above, wherein the 4- to 7-memberednitrogenous saturated heterocycle is an azetidine, pyrrolidine, orpiperidine ring;

[6] the compound according to [1] above, represented by general formula(I-1)

(in the formula, ring1-1 represents a benzene, cyclohexane, or pyridinering, each of which may be substituted by from one to five substituentseach independently selected from the group consisting of (1) halogenatoms, (2) C₁₋₄ alkyl groups, (3) C₁₋₄ alkoxy groups, (4) nitrile, and(5) CF₃, and ring2-1 represents a 4- to 7-membered nitrogenous saturatedheterocycle, which may be substituted by from one to three —K—R²,wherein the other symbols have the same definitions as above);

[7] the compound according to [6] above, wherein R² is a C₂₋₄ alkenylgroup or a C₂₋₄ alkynyl group, each of which may be substituted by fromone to five substituents each independently selected from the groupconsisting of (1) NR³R⁴, (2) halogen atoms, (3) CONR⁵R⁶, (4) CO₂R⁷, and(5) OR⁸;

[8] the compound according to [1] above, which is (1)9-(1-acryloyl-3-azetidinyl)-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,(2)6-amino-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,(3)9-[(l-acryloyl-4-piperidinyl)methyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,(4)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,(5)6-amino-9-{(3S)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,(6)6-amino-7-[4-(3-chlorophenoxy)phenyl]-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7,9-dihydro-8H-purin-8-one,(7)6-amino-9-[1-(2-butynoyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,or (8)6-amino-9-{1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one,or an optical isomer thereof or their mixture;

[9] a pharmaceutical composition comprising a compound represented bygeneral formula (I) according to [1] above, an optical isomer thereof ortheir mixture, a salt thereof, a solvate thereof, an N-oxide thereof, ora prodrug thereof;

[10] the pharmaceutical composition according to [9] above, that is aBtk inhibitor;

[11] the pharmaceutical composition according to [9]above, that is aprophylactic agent and/or a therapeutic agent for a Btk-related disease;

[12] the pharmaceutical composition according to [11] above, wherein theBtk-related disease is an allergic disease, an autoimmune disease, aninflammatory disease, a thromboembolic disease, or a cancer;

[13] the pharmaceutical composition according to [12] above, wherein thecancer is a non-Hodgkin's lymphoma;

[14] the pharmaceutical composition according to [9] above, which is aninhibitor of B-cell activation;

[15] a method of preventing and/or treating a Btk-related disease,comprising administering to a mammal of an effective amount of acompound represented by general formula (I) according to [1] above, anoptical isomer thereof or their mixture, a salt thereof, a solvatethereof, an N-oxide thereof, or a prodrug thereof;

[16] a compound represented by general formula (I) according to [1]above, an optical isomer thereof or their mixture, a salt thereof, asolvate thereof, an N-oxide thereof, or a prodrug thereof, forpreventing and/or treating a Btk-related disease; and

[17] use of a compound represented by general formula (I) according to[1] above, an optical isomer thereof or their mixture, a salt thereof, asolvate thereof, an N-oxide thereof, or a prodrug thereof, to produce anagent for preventing and/or treating a Btk-related disease.

The compounds of the present invention, in addition to having aBtk-selective inhibitory activity, exhibit an excellent metabolicstability and can avoid hepatotoxicity or the like, and as a consequenceare useful as very safe therapeutic agents for diseases in which B cellsand/or mast cells participate, such as non-Hodgkin's lymphoma.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention is described in detail below.

In the present invention, “having a Btk-selective inhibitory activity”denotes having a Btk-selective inhibitory activity with respect tonon-Btk tyrosine kinases and particularly lymphocyte-specific proteintyrosine kinase (Lck), protein-tyrosine kinase fyn (Fyn), and v-yes-1Yamaguchi sarcoma viral-related oncogene homolog isoform A (LynA). Thisproperty makes it possible to avoid unpredictable adverse reactionscaused by the inhibition of other tyrosine kinases. For example, theappearance of retinal abnormalities is known in Lck-deficient mice(Oncogene, Volume 16, pp. 2351-2356, 1998), which raises the possibilitythat adverse reactions will be caused in the eye in the event of aninhibition of Lck.

In the present invention, the halogen atom denotes fluorine, chlorine,bromine, and iodine.

In the present invention, the C₁₋₄ alkyl group denotes straight-chainand branched-chain C₁₋₄ alkyl groups, e.g., methyl, ethyl, propyl,isopropyl, butyl, sec-butyl, and tert-butyl.

In the present invention, the C₁₋₄ alkylene group denotes methylene,ethylene, propylene, butylene, and their isomers.

In the present invention, the C₁₋₄ alkoxy group denotes straight-chainand branched-chain C₁₋₄ alkoxy groups, e.g., methoxy, ethoxy, propoxy,isopropoxy, butoxy, isobutyloxy, and tert-butoxy.

In the present invention, the C₂₋₄ alkenyl group denotes straight-chainand branched-chain C₂₋₄ alkenyl groups, e.g., ethenyl, 1-propenyl,2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, and 1,3-butadienyl.

In the present invention, the C₂₋₄ alkynyl group denotes straight-chainand branched-chain C₂₋₄ alkynyl groups, e.g., ethynyl, 1-propynyl,2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, and 1,3-butadiynyl.

In the present invention, the C₁₋₄ haloalkyl group denotes a groupprovided by substituting one or two or more halogen atoms into a C₁₋₄alkyl group, and can be exemplified by a fluoromethyl group, achloromethyl group, a bromomethyl group, an iodomethyl group, adifluoromethyl group, a trifluoromethyl group, a 1-fluoroethyl group, a2-fluoroethyl group, a 2-chloroethyl group, a pentafluoroethyl group, a1-fluoropropyl group, a 2-chloropropyl group, a 3-fluoropropyl group, a3-chloropropyl group, a 4,4,4-trifluorobutyl group, and a 4-bromobutylgroup.

In the present invention, the C₁₋₄ haloalkoxy group denotes a groupprovided by substituting one or two or more halogen atoms into a C₁₋₄alkoxy group, and can be exemplified by a trifluoromethoxy group, atrichloromethoxy group, a chloromethoxy group, a bromomethoxy group, afluoromethoxy group, an iodomethoxy group, a difluoromethoxy group, adibromomethoxy group, a 2-chloroethoxy group, a 2,2,2-trifluoroethoxygroup, a 2,2,2-trichloroethoxy group, a 3-bromopropoxy group, a3-chloropropoxy group, a 2,3-dichloropropoxy group, a 1-fluorobutoxygroup, a 4-fluorobutoxy group, and a 1-chlorobutoxy group.

In the present invention, the 4- to 7-membered cyclic group denotes aC₄₋₇ carbocyclic ring or a 4- to 7-membered heterocycle.

In the present invention, the C₄₋₇ carbocyclic ring denotes a C₄₋₇monocyclic aliphatic or aromatic carbocyclic ring. The aliphatic systemmay be partially or completely saturated. Examples here are cyclobutane,cyclopentane, cyclohexane, cycloheptane, cyclopentene, cyclohexene,cycloheptene, cyclobutadiene, cyclopentadiene, cyclohexadiene,cycloheptadiene, and benzene.

In the present invention, the C₅₋₆ carbocyclic ring denotes a C₅₋₆monocyclic aliphatic or aromatic carbocyclic ring. The aliphatic systemmay be partially or completely saturated. Examples here arecyclopentane, cyclohexane, cyclopentene, cyclohexene, cyclopentadiene,cyclohexadiene, and benzene.

In the present invention, the 4- to 7-membered heterocycle denotes a 4-to 7-membered unsaturated heterocycle or a 4- to 7-membered saturatedheterocycle.

In the present invention, the 4- to 7-membered unsaturated heterocycledenotes an unsaturated 4- to 7-membered monocyclic heterocycle thatcontains from one to five hetero atoms selected from the oxygen atom,nitrogen atom, and sulfur atom, and can be exemplified by pyrrole,imidazole, triazole, tetrazole, pyrazole, pyridine, pyrazine,pyrimidine, pyridazine, azepine, diazepine, furan, pyran, oxepine,thiophene, thiopyran, thiepine, oxazole, isoxazole, thiazole,isothiazole, furazan, oxadiazole, oxazine, oxadiazine, oxazepine,oxadiazepine, thiadiazole, thiazine, thiadiazine, thiazepine, andthiadiazepine.

In the present invention, the 4- to 7-membered saturated heterocycledenotes a partially or completely saturated 4- to 7-membered monocyclicheterocycle that contains from one to five hetero atoms eachindependently selected from the oxygen atom, nitrogen atom, and sulfuratom, and can be exemplified by azetidine, pyrroline, pyrrolidine,imidazoline, imidazolidine, triazoline, triazolidine, tetrazoline,tetrazolidine, pyrazoline, pyrazolidine, dihydropyridine,tetrahydropyridine, piperidine, dihydropyrazine, tetrahydropyrazine,piperazine, dihydropyrimidine, tetrahydropyrimidine, perhydropyrimidine,dihydropyridazine, tetrahydropyridazine, perhydropyridazine,dihydroazepine, tetrahydroazepine, perhydroazepine, dihydrodiazepine,tetrahydrodiazepine, perhydrodiazepine, oxetane, dihydrofuran,tetrahydrofuran, dihydropyran, tetrahydropyran, dihydrooxepine,tetrahydrooxepine, perhydrooxepine, thietane, dihydrothiophene,tetrahydrothiophene, dihydrothiopyran, tetrahydrothiopyran,dihydrothiepine, tetrahydrothiepine, perhydrothiepine, dihydrooxazole,tetrahydrooxazole (oxazolidine), dihydroisoxazole, tetrahydroisoxazole(isoxazolidine), dihydrothiazole, tetrahydrothiazole (thiazolidine),dihydroisothiazole, tetrahydroisothiazole (isothiazolidine),dihydrofurazan, tetrahydrofurazan, dihydrooxadiazole,tetrahydrooxadiazole (oxadiazolidine), dihydrooxazine,tetrahydrooxazine, dihydrooxadiazine, tetrahydrooxadiazine,dihydrooxazepine, tetrahydrooxazepine, perhydrooxazepine,dihydrooxadiazepine, tetrahydrooxadiazepine, perhydrooxadiazepine,dihydrothiadiazole, tetrahydrothiadiazole (thiadiazolidine),dihydrothiazine, tetrahydrothiazine, dihydrothiadiazine,tetrahydrothiadiazine, dihydrothiazepine, tetrahydrothiazepine,perhydrothiazepine, dihydrothiadiazepine, tetrahydrothiadiazepine,perhydrothiadiazepine, morpholine, thiomorpholine, oxathiane, dioxolane,dioxane, dithiolane, and dithiane.

In the present invention, the 4- to 7-membered nitrogenous saturatedheterocycle refers to those 4- to 7-membered saturated heterocycles thatnecessarily contain at least one nitrogen atom. Examples are azetidine,pyrroline, pyrrolidine, imidazoline, imidazolidine, triazoline,triazolidine, tetrazoline, tetrazolidine, pyrazoline, pyrazolidine,dihydropyridine, tetrahydropyridine, piperidine, dihydropyrazine,tetrahydropyrazine, piperazine, dihydropyrimidine, tetrahydropyrimidine,perhydropyrimidine, dihydropyridazine, tetrahydropyridazine,perhydropyridazine, dihydroazepine, tetrahydroazepine, perhydroazepine,dihydrodiazepine, tetrahydrodiazepine, perhydrodiazepine,dihydrooxazole, tetrahydrooxazole (oxazolidine), dihydroisoxazole,tetrahydroisoxazole (isoxazolidine), dihydrothiazole, tetrahydrothiazole(thiazolidine), dihydroisothiazole, tetrahydroisothiazole(isothiazolidine), dihydrofurazan, tetrahydrofurazan, dihydrooxadiazole,tetrahydrooxadiazole (oxadiazolidine), dihydrooxazine,tetrahydrooxazine, dihydrooxadiazine, tetrahydrooxadiazine,dihydrooxazepine, tetrahydrooxazepine, perhydrooxazepine,dihydrooxadiazepine, tetrahydrooxadiazepine, perhydrooxadiazepine,dihydrothiadiazole, tetrahydrothiadiazole (thiadiazolidine),dihydrothiazine, tetrahydrothiazine, dihydrothiadiazine,tetrahydrothiadiazine, dihydrothiazepine, tetrahydrothiazepine,perhydrothiazepine, dihydrothiadiazepine, tetrahydrothiadiazepine,perhydrothiadiazepine, morpholine, and thiomorpholine.

The 4- to 6-membered nitrogenous saturated heterocycles can beexemplified in the present invention by azetidine, pyrroline,pyrrolidine, imidazoline, imidazolidine, triazoline, triazolidine,tetrazoline, tetrazolidine, pyrazoline, pyrazolidine, dihydropyridine,tetrahydropyridine, piperidine, dihydropyrazine, tetrahydropyrazine,piperazine, dihydropyrimidine, tetrahydropyrimidine, perhydropyrimidine,dihydropyridazine, tetrahydropyridazine, perhydropyridazine,dihydrooxazole, tetrahydrooxazole (oxazolidine), dihydroisoxazole,tetrahydroisoxazole (isoxazolidine), dihydrothiazole, tetrahydrothiazole(thiazolidine), dihydroisothiazole, tetrahydroisothiazole(isothiazolidine), dihydrofurazan, tetrahydrofurazan, dihydrooxadiazole,tetrahydrooxadiazole (oxadiazolidine), dihydrooxazine,tetrahydrooxazine, dihydrooxadiazine, tetrahydrooxadiazine,dihydrothiadiazole, tetrahydrothiadiazole (thiadiazolidine),dihydrothiazine, tetrahydrothiazine, dihydrothiadiazine,tetrahydrothiadiazine, morpholine, and thiomorpholine.

L in the present invention is preferably —O—, —CH₂—O—, —O—CH₂—, —NH—,—C(O)—, —CH₂—, or —CH(OH)—.

R¹ in the present invention is preferably a halogen atom or a C₁₋₄alkoxy group.

The 4- to 7-membered cyclic group for ring1 in the present invention ispreferably a C₅₋₆ carbocyclic ring or a 4- to 7-membered unsaturatedheterocycle and is more preferably a benzene, cyclohexane, or pyridinering.

The 4- to 7-membered saturated heterocycle for ring2 in the presentinvention is preferably a 4- to 7-membered nitrogenous saturatedheterocycle, is more preferably a 4- to 6-membered nitrogenous saturatedheterocycle, is even more preferably an azetidine, pyrrolidine, orpiperidine ring, and particularly preferably is an azetidine orpyrrolidine ring.

K in the present invention is preferably a bond or —C(O)—.

R² in the present invention is preferably a C₂₋₄ alkenyl group or a C₂₋₄alkynyl group, each of which may be substituted by from one to fivesubstituents each independently selected from the group consisting of(1) NR³R⁴, (2) halogen atoms, (3) CONR⁵R⁶, (4) CO₂R⁷, and (5) OR⁸.

m in the present invention is preferably 0 or 1 and more preferably is0.

Any combination of the individual preferred groups provided as examplesabove and selected from L, R¹, ring1, ring2, K, R², and m is alsopreferred in the present invention.

Compounds represented by general formula (I-1)

(all of the symbols in the formula have the same definitions as above)are preferred in the present invention among compounds with generalformula (I).

R¹ in general formula (I-1) is preferably a halogen atom or a C₁₋₄alkoxy group.

The ring1-1 in general formula (I-1) is preferably a benzene ring.

The 4- to 7-membered nitrogenous saturated heterocycle for ring2-1 ingeneral formula (I-1) is preferably an azetidine, pyrrolidine, orpiperidine ring and more preferably is an azetidine or pyrrolidine ring.

With regard to the —K—R²— substituent on the ring2-1 in general formula(I-1), K is preferably a bond or —C(O)— and R² is preferably a C₂₋₄alkenyl group or C₂₋₄ alkynyl group, each of which may be substituted byfrom one to five substituents each independently selected from the groupconsisting of (1) NR³R⁴, (2) halogen atoms, (3) CONR⁵R⁶, (4) CO₂R⁷, and(5) OR⁸.

m in general formula (I-1) is preferably 0 or 1 and more preferably is0.

Any combination of the individual preferred groups provided as examplesabove is also preferred for general formula (I-1).

[Isomers]

The present invention encompasses all isomers, unless specificallyindicated otherwise. For example, the alkyl groups include bothstraight-chain alkyl groups and branched-chain alkyl groups. Moreover,all of the following are included in the present invention: geometricisomers (E configuration, Z configuration, cis configuration, transconfiguration) for double bonds, rings, and condensed rings; opticalisomers due to, for example, the presence of an asymmetric carbon atom(R and S configurations, α and β positions, enantiomers, diastereomers);optically active forms that exhibit optical rotation (D, L, d, and lconfigurations); polar forms generated by chromatographic separation(high-polarity forms, low-polarity forms); equilibrium compounds;rotational isomers; mixtures of the preceding in any proportions; andracemic mixtures. The present invention also encompasses all isomersarising due to tautomers.

In addition, an optical isomer in the present invention refers not onlyto the 100% pure optical isomer, but may also include another opticalisomer at less than 50%.

In the present invention, unless specifically stated otherwise, and asis clear to the individual skilled in the art, the

[C 11]

symbol represents bonding toward the back side of the plane of the paper(that is, the α-position); the

[C 12]

symbol represents bonding toward the front side of the plane of thepaper (that is, the β-position); and

[C 13]

represents the α-position, β-position, or their mixture in anyproportion.

The compounds represented by general formula (I) are converted into thecorresponding salts by known methods. The salt is preferably awater-soluble salt. Suitable salts can be exemplified by the salts withalkali metals (potassium, sodium, and so forth), salts withalkaline-earth metals (calcium, magnesium, and so forth), the ammoniumsalt, salts with pharmaceutically acceptable organic amines(tetramethylammonium, triethylamine, methylamine, dimethylamine,cyclopentylamine, benzylamine, phenethylamine, piperidine,monoethanolamine, diethanolamine, tris(hydroxymethyl)aminomethane,lysine, arginine, N-methyl-D-glucamine, and so forth), and acid additionsalts (inorganic acid salts (the hydrochloride, hydrobromide,hydroiodide, sulfate, phosphate, nitrate, and so forth) and organic acidsalts (the acetate, trifluoroacetate, lactate, tartrate, oxalate,fumarate, maleate, benzoate, citrate, methanesulfonate, ethanesulfonate,benzenesulfonate, toluenesulfonate, isethionate, glucuronate, gluconate,and so forth)).

The compounds represented by general formula (I) and their salts canalso be converted into solvates. This solvate preferably is watersoluble and has a low toxicity. Suitable solvates can be exemplified bysolvates with, for example, water or an alcohol solvent (for example,ethanol).

A prodrug of a compound represented by general formula (I) denotes acompound that is converted by a reaction in vivo, e.g., by an enzyme orgastric acid, into the compound represented by general formula (I).Prodrugs of the compounds with general formula (I) can be exemplified bycompounds in which—when the compound represented by general formula (I)has a hydroxyl group—this hydroxyl group is acylated, alkylated,phosphated, or borated (for example, compounds provided by theacetylation, palmitoylation, propanoylation, pivaloylation,succinylation, fumarylation, alanylation, ordimethylaminomethylcarbonylation of the hydroxyl group in the compoundsof the invention); other examples are compounds provided by theesterification or amidation of a carboxyl group in a compoundrepresented by general formula (I) (for example, compounds provided bythe ethyl esterification, isopropyl esterification, phenylesterification, carboxymethyl esterification, dimethylaminomethylesterification, pivaloyloxymethyl esterification, ethoxycarbonyloxyethylesterification, phthalidyl esterification,(5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl esterification,cyclohexyloxycarbonylethyl esterification, or methylamidation of acarboxyl group in a compound represented by general formula (I)). Thesecompounds can be prepared by known methods. In addition, the prodrug ofa compound represented by general formula (I) may be a hydrate oranhydrous. The prodrug of a compound represented by general formula (I)is converted under physiological conditions into the compoundrepresented by general formula (I), as described on pages 163 to 198 ofMolecular Design in Volume 7 of Development of Medicines (HirokawaShoten Co., 1990). In addition, the compounds represented by generalformula (I) may be labeled with an isotope (for example, ²H, ³H, ¹¹C,¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³⁵S, ¹⁸F, ³⁶Cl, ¹²³I, and ¹²⁵I)

[Methods for Producing the Compounds of the Present Invention]

The compounds of the present invention can be produced by suitablymodifying and combining known methods, for example, the methodsdescribed in Comprehensive Organic Transformations: A Guide toFunctional Group Preparations, 2nd Edition (Richard C. Larock, JohnWiley & Sons Inc., 1999), or the methods given in the examples.

Among the compounds represented by general formula (I), compounds inwhich the ring2 is an unsubstituted 4- to 7-membered nitrogenoussaturated heterocycle, i.e., compounds represented by general formula(I-A)

(in the formula, ring2^(a) represents an unsubstituted 4- to 7-memberednitrogenous saturated heterocycle, while the other symbols have the samedefinitions as above), can be produced using the following reactionscheme 1.

(In the formulas, T¹ and T² each independently represent a protectivegroup for the amino group (for example, the benzyl (Bn) group,4-methoxybenzyl group, and 3,4-dimethoxybenzyl group); X¹ and X² eachindependently represent a halogen atom; ring2-1 represents a ring2^(a)that has been protected by a protective group represented by T³ (forexample, the tert-butoxycarbonyl (Boc) group and benzyloxycarbonyl (Cbz)group); and the other symbols have the same definitions as above.)

The reaction 1 in reaction scheme 1 is known and is carried out using acompound with general formula (a) and a protected amine derivative,i.e., a compound represented by T¹T²-NH (T¹ and T² in the formula havethe same definitions as above) and running the reaction in an organicsolvent (for example, dichloromethane, tetrahydrofuran, dioxane,dimethylformamide, dimethylacetamide, and 1-methyl-2-pyrrolidone) in thepresence of a base (for example, triethylamine,N,N-diisopropylethylamine, pyridine, 4-dimethylaminopyridine, andN-methylmorpholine) at a temperature from −20° C. to room temperature.

The reaction 2 in reaction scheme 1 is known and is carried out using acompound with general formula (b) and a compound represented by generalformula (II) and running the reaction in an organic solvent (forexample, dioxane, acetonitrile, tetrahydrofuran, dimethylformamide,dimethylacetamide, and 1-methyl-2-pyrrolidone) in the presence of a base(for example, triethylamine, N,N-diisopropylethylamine, pyridine,4-dimethylaminopyridine, and N-methylmorpholine) at a temperature from−20° C. to 70° C.

The reaction 3 in reaction scheme 1 is known and is carried out using acompound represented by general formula (c) and using a metal reagent(for example, zinc, iron, tin, tin chloride, iron chloride, samarium,indium, and sodium borohydride-nickel chloride) in a water-misciblesolvent (for example, ethanol, methanol, tetrahydrofuran, and ethylacetate) in the presence or absence of an acid (for example,hydrochloric acid, hydrobromic acid, ammonium chloride, acetic acid, andammonium formate) at a temperature of 0° C. to 150° C.

The reaction 4 in reaction scheme 1 is known and is carried out using acompound represented by general formula (d) and using a reagent (forexample, 1,1′-carbonyldiimidazole (CDI) and triphosgene) in an organicsolvent (for example, tetrahydrofuran, dimethylformamide, anddimethylacetamide) in the presence or absence of a base (for example,triethylamine, N,N-diisopropylethylamine, pyridine,4-dimethylaminopyridine, and N-methylmorpholine) at from a temperaturegenerated by ice cooling to the reflux temperature.

The deprotection reactions for the protective groups in reaction scheme1 are known and can be run by the methods described below. Examples hereare (1) deprotection reactions based on alkaline hydrolysis, (2)deprotection reactions under acidic conditions, (3) deprotectionreactions based on hydrogenolysis, (4) deprotection reactions for thesilyl group, (5) deprotection reactions using a metal, and (6)deprotection reactions using a metal complex.

These methods are specifically described below.

(1) The deprotection reaction based on alkaline hydrolysis may becarried out, for example, in an organic solvent (for example, methanol,tetrahydrofuran, and dioxane) at from 0° C. to 40° C. using an alkalimetal hydroxide (for example, sodium hydroxide, potassium hydroxide, andlithium hydroxide), an alkaline-earth metal hydroxide (for example,barium hydroxide and calcium hydroxide), or a carbonate (for example,sodium carbonate and potassium carbonate), or an aqueous solution of thepreceding, or their mixture.

(2) The deprotection reaction under acidic reactions may be carried out,for example, at from 0° C. to 100° C. in the presence or absence of2,2,2-trifluoroethanol, in an organic solvent (for example,dichloromethane, chloroform, dioxane, ethyl acetate, methanol, isopropylalcohol, tetrahydrofuran, and anisole) and in an organic acid (forexample, acetic acid, trifluoroacetic acid, methanesulfonic acid, andp-tosylate) or an inorganic acid (for example, hydrochloric acid andsulfuric acid) or their mixture (for example, hydrogen bromide/aceticacid).

(3) The deprotection reaction based on hydrogenolysis may be carriedout, for example, at 0° C. to 200° C. in a solvent (for example, anether (for example, tetrahydrofuran, dioxane, dimethoxyethane, anddiethyl ether), an alcohol (for example, methanol and ethanol), abenzene solvent (for example, benzene and toluene), a ketone (forexample, acetone and methyl ethyl ketone), a nitrile (for example,acetonitrile), an amide (for example, N,N-dimethylformamide), water,ethyl acetate, acetic acid, or a mixed solvent of two or more of thepreceding) in the presence of a catalyst (for example, palladium-carbon,palladium black, palladium hydroxide-carbon, platinum oxide, and Raneynickel) in a hydrogen atmosphere at normal pressure or elevated pressureor in the presence of ammonium formate.

(4) The deprotection reaction for the silyl group may be carried out,for example, at 0° C. to 40° C. in a water-miscible organic solvent (forexample, tetrahydrofuran and acetonitrile) using tetrabutylammoniumfluoride. It may also be carried out, for example, at −10° C. to 100° C.in an organic acid (for example, acetic acid, trifluoroacetic acid,methanesulfonic acid, and p-tosylate) or an inorganic acid (for example,hydrochloric acid and sulfuric acid) or their mixture (for example,hydrogen bromide/acetic acid).

(5) The deprotection reaction using a metal may be carried out, forexample, at 0° C. to 40° C. in an acidic solvent (for example, aceticacid, a buffer with a pH of 4.2 to 7.2, or a mixed solution of thepreceding solutions with an organic solvent such as tetrahydrofuran) inthe presence of zinc powder and as necessary while applying ultrasound.

(6) The deprotection reaction using a metal complex may be carried out,for example, at 0° C. to 40° C. in an organic solvent (for example,dichloromethane, N,N-dimethylformamide, tetrahydrofuran, ethyl acetate,acetonitrile, dioxane, and ethanol), water, or their mixed solutions,using a metal complex (for example,tetrakis(triphenylphosphine)palladium(0),bis(triphenylphosphine)palladium(II) dichloride, palladium(II) acetate,and tris(triphenylphosphine)rhodium(I) chloride) in the presence of atrapping reagent (for example, tributyltin hydride, triethylsilane,dimedone, morpholine, diethylamine, and pyrrolidone), an organic acid(for example, acetic acid, formic acid, and 2-ethylhexanoic acid),and/or an organic acid salt (for example, sodium 2-ethylhexanoate andpotassium 2-ethylhexanoate) and in the presence or absence of aphosphine-type reagent (for example, triphenylphosphine).

In addition to the preceding, the deprotection reaction may also becarried out using the methods described in, for example, T. W. Greene,Protective Groups in Organic Synthesis, Wiley, New York, 1999.

The protective group for an amino group can be exemplified by thebenzyloxycarbonyl group, t-butoxycarbonyl group, allyloxycarbonyl(Alloc) group, 1-methyl-1-(4-biphenyl)ethoxycarbonyl (Bpoc) group,trifluoroacetyl group, 9-fluorenylmethoxycarbonyl group, benzyl (Bn)group, p-methoxybenzyl group, benzyloxymethyl (BOM) group, and2-(trimethylsilyl)ethoxymethyl (SEM) group.

In addition to the preceding, the protective group for an amino groupmay be any group capable of a facile and selective elimination and isnot otherwise particularly limited. For example, the groups described inT. W. Greene, Protective Groups in Organic Synthesis, Wiley, New York,1999, may be used.

The reaction 5 in reaction scheme 1 is known and is carried out using acompound with general formula (f) and a compound represented by generalformula (III-1) or general formula (III-2) and running the reaction atfrom room temperature to 120° C. in an organic solvent (for example,dichloromethane and acetonitrile) in the presence of a base (forexample, pyridine, triethylamine, and N,N-diisopropylethylamine), acopper salt (for example, copper(II) acetate), and a drying agent (forexample, molecular sieve).

Among compounds with general formula (I), compounds in which ring2 is a4- to 7-membered nitrogenous saturated heterocycle that is substitutedby at least one —K—R² wherein K represents C(O) and R² represents a C₁₋₄alkylene or C₂₋₄ alkenylene group which may be substituted by halogen,i.e., compounds represented by general formula (I-B)

(in the formula, ring2^(b) represents a 4- to 7-membered nitrogenoussaturated heterocycle substituted by —K^(b)—R^(2b); K^(b) represents—C(O)—; R^(2b) represents a C₁₋₄ alkylene or C₂₋₄ alkenylene group whichmay be substituted by halogen; and the other symbols have the samedefinitions as above), can be produced using a compound with generalformula (I-A), which can be produced by reaction scheme 1, and acompound represented by general formula (I-B-1)[C 17]X^(b)—C(O)—R^(2b)  (I-B-1)(in the formula, X^(b) represents a halogen atom and R^(2b) has the samedefinition as above) and carrying out a reaction at a temperature from0° C. to room temperature in an organic solvent (for example,dichloromethane, chloroform, dimethylformamide, dimethylacetamide,diethyl ether, and tetrahydrofuran) in the presence of a base (forexample, triethylamine, pyridine, N,N-diisopropylethylamine,4-dimethylaminopyridine, and N-methylmorpholine).

Among compounds with general formula (I), compounds in which ring2 is a4- to 7-membered nitrogenous saturated heterocycle substituted by atleast one —K—R² wherein K represents C(O) and R² represents a C₁₋₄alkylene or C₂₋₄ alkenylene group which may be substituted by NR³R⁴,i.e., compounds with general formula (I-C)

(in the formula, ring2^(c) represents a 4- to 7-membered nitrogenoussaturated heterocycle substituted by —K^(b)—R^(2c);R^(2c) represents a C₁₋₄ alkylene or C₂₋₄ alkenylene group which may besubstituted by NR³R⁴; and the other symbols have the same definitions asabove), can be produced by reacting a compound represented by generalformula (I-C-2)

(all of the symbols in the formula are defined as above) at from roomtemperature to 120° C. in an organic solvent (for example,tetrahydrofuran, dioxane, acetonitrile, dimethylformamide,dimethylacetamide, and 1-methyl-2-pyrrolidone) with the compoundobtained by reacting a compound represented by general formula (I-A)with a compound represented by general formula (I-C-1)[C 19]X^(c-1)—C(O)—R^(2c-1)—X^(c-2)  (I-C-1)(in the formula, X^(c-1) and X^(c-2) each independently represent ahalogen atom and R^(2c-1) represents a C₁₋₄ alkylene or C₂₋₄ alkenylenegroup) at or below 0° C. in an organic solvent (for example,dichloromethane, chloroform, dimethylformamide, dimethylacetamide,diethyl ether, and tetrahydrofuran) in the presence of a base (forexample, triethylamine, pyridine, N,N-diisopropylethylamine,4-dimethylaminopyridine, and N-methylmorpholine).

Among compounds with general formula (I), compounds in which ring2 is a4- to 7-membered nitrogenous saturated heterocycle substituted by atleast one —K—R² wherein K represents C(O) and R² represents a C₁₋₄alkylene, C₂₋₄ alkenylene, or C₂₋₄ alkynylene group which may besubstituted by CONR⁵R⁶, CO₂R⁷, or OR⁸, i.e., compounds with generalformula (I-D)

(in the formula, ring2^(d) represents a 4- to 7-membered nitrogenoussaturated heterocycle substituted by at least one —K^(b)—R^(2d); R^(2d)represents a C₁₋₄ alkylene, C₂₋₄ alkenylene, or C₂₋₄ alkynylene groupwhich may be substituted by CONR⁵R⁶, CO₂R⁷, or OR⁸; and the othersymbols have the same definitions as above), can be produced using acompound represented by general formula (I-A) and a compound representedby general formula (I-D-1)[C 22]R^(2d)—CO₂H  (I-D-1)(R^(2d) in the formula is defined as above) and carrying out a reactionat from 0° C. to room temperature using a condensing agent (for example,1,3-dicyclohexylcarbodiimide (DCC),1-ethyl-3-[3-(dimethylamino)propyl]carbodiimide (EDC),1,1′-carbonyldiimidazole (CDI), 2-chloro-1-methylpyridinium iodide, and1-propanephosphonic acid cyclic anhydride (PPA)) in an organic solvent(for example, chloroform, dichloromethane, dimethylformamide,dimethylacetamide, diethyl ether, and tetrahydrofuran), or without asolvent, in the presence or absence of a base (for example,triethylamine, pyridine, N,N-diisopropylethylamine,4-dimethylaminopyridine, and N-methylmorpholine) and with or without theuse of 1-hydroxybenztriazole (HOBt).

The compounds used as starting materials in each of the reactions inthis Description, for example, general formula (a), (II), (III-1),(III-2), (I-B-1), (I-C-1), (I-C-2), and (I-D-1), are either known or canbe readily produced by known methods.

For each of the reactions in this Description, the reactions accompaniedby the application of heat can be carried out using a water bath, oilbath, sand bath, or microwaves, as will be clear to the individualskilled in the art.

A solid-phase reagent, comprising the reagent carried on a highmolecular weight polymer (for example, polystyrene, polyacrylamide,polypropylene, and polyethylene glycol), may be used as appropriate inthe reactions in this Description.

The reaction product in each of the reactions of this Description can bepurified by the usual purification means, for example, using methodssuch as distillation at normal pressure or under reduced pressure,high-performance liquid chromatography using silica gel or magnesiumsilicate, thin-layer chromatography, ion-exchange resins, scavengerresins, column chromatography, washing, recrystallization, and so forth.Purification may be carried out in each reaction or may be performedafter the completion of several reactions.

[Toxicity]

The compounds of the present invention have an acceptably low toxicity,for example, they have almost no CYP inhibitory activity and almost nohepatotoxicity, and thus can be safely used as an active pharmaceuticalingredient.

[Application to Pharmaceuticals]

The compounds of the present invention have a selective Btk inhibitoryactivity and as a result are useful as agents for preventing and/ortreating Btk-related diseases, i.e., diseases in which B cells and/ormast cells participate, for example, allergic diseases, autoimmunediseases, inflammatory diseases, thromboembolic diseases, cancers, andgraft-versus-host diseases. The compounds of the present invention alsoexercise a selective inhibitory action on B cell activation and as aresult are also effective as inhibitors of B cell activation.

The allergic diseases in the present invention can be exemplified byallergies, anaphylaxis, allergic conjunctivitis, allergic rhinitis, andallergic dermatitis.

The autoimmune diseases in the present invention can be exemplified byinflammatory bowel disease, arthritis, lupus, rheumatoid arthritis,psoriatic arthritis, osteoarthritis, Still's disease, juvenilearthritis, type I diabetes, myasthenia gravis, Hashimoto's thyroiditis,Ord's thyroiditis, Basedow's disease, Sjögren's syndrome, multiplesclerosis, Guillain-Barré syndrome, acute disseminatedencephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome,ankylosing spondylitis, anti-phospholipid antibody syndrome, aplasticanemia, autoimmune hepatitis, celiac disease, Goodpasture's syndrome,idiopathic thrombocytopenic purpura, optic neuritis, scleroderma,primary biliary cirrhosis, Reiter's disease, Takayasu's arteritis,temporal arteritis, warm autoimmune hemolytic anemia, Wegener'sgranuloma, psoriasis, alopecia universalis, Behçet's disease, chronicfatigue syndrome, dysautonomia, endometriosis, interstitial cystitis,myotonia, vulvodynia, and systemic lupus erythematosus.

The inflammatory diseases in the present invention can be exemplified byasthma, appendicitis, blepharitis, bronchiolitis, bronchitis, bursitis,cervicitis, cholangitis, cholecystitis, colitis, conjunctivitis,cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis,endocarditis, endometritis, enteritis, epicondylitis, epididymitis,fasciitis, fibrositis, gastritis, gastroenteritis, hepatitis,hidradenitis suppurativa, laryngitis, mastitis, meningitis, myelitis,myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis,pancreatitis, parotitis, pericarditis, peritonitis, pharyngitis,pleuritis, phlebitis, pneumonia, proctitis, prostatitis, pyelonephritis,rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis,tonsillitis, uveitis, vaginitis, vasculitis, and vulvitis.

The thromboembolic diseases in the present invention can be exemplifiedby myocardial infarction, angina pectoris, reocclusion afterangioplasty, restenosis after angioplasty, reocclusion afteraortocoronary bypass, restenosis after aortocoronary bypass, stroke,transient ischemia, peripheral arterial occlusive disorders, pulmonaryembolism, and deep venous thrombosis.

In the present invention, the cancers include non-Hodgkin's lymphomas,among which B-cell non-Hodgkin's lymphomas are particularly suitable,for example, Burkitt's lymphoma, AIDS-related lymphoma, marginal zoneB-cell lymphoma (nodal marginal zone B-cell lymphoma, extranodalmarginal zone B-cell lymphoma, and splenic marginal zone B-celllymphoma), diffuse large B-cell lymphoma, primary effusion lymphoma,lymphomatoid granulomatosis, follicular lymphoma, B-cell chroniclymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacyticleukemia/Waldenstrom's macroglobulinemia, plasmacytoma, mantle celllymphoma, mediastinal large B-cell lymphoma, intravascular large B-celllymphoma, and hairy cell leukemia. In addition to non-Hodgkin'slymphoma, the cancers in the present invention include pancreaticendocrine tumors, for example, insulinoma, gastrinoma, glucagonoma,somatostatinoma, VIPoma, PPoma, and GRFoma.

A compound of the present invention may be administered by itself or maybe administered as a combined preparation combined with another drug forthe purpose of

1) supplementing and/or enhancing the prophylactic and/or therapeuticeffect of the compound,

2) improving the pharmacokinetics and absorption and reducing the doseof the compound, and/or

3) reducing the adverse reactions of the compound.

The combined preparation of a compound of the present invention withanother drug may be administered in the form of a compounded agent inwhich both components are compounded in a single formulation or may beadministered as separate formulations. Administration as separateformulations includes simultaneous administration and administration atdifferent times. In the case of administration at different times, thecompound of the present invention may be administered first followed byadministration of the other drug, or the other drug may be administeredfirst followed by the administration of the compound of the presentinvention. The same method of administration may be used for each ordifferent methods of administration may be used.

There is no particular limitation on the diseases that may be subjectedto the prevention and/or treatment by the aforementioned combinedpreparation, and this combined preparation may be used with any diseasewith which the prevention and/or treatment effect of the compound of thepresent invention is supplemented and/or enhanced.

This other drug for supplementing and/or enhancing the prophylacticand/or therapeutic effect of the compounds of the present inventionagainst allergic diseases can be exemplified by antihistamines,leukotriene antagonists, antiallergic drugs, thromboxane A2 receptorantagonists, thromboxane synthase inhibitors, and steroids.

The other drug for supplementing and/or enhancing the prophylacticand/or therapeutic effect of the compounds of the present inventionagainst autoimmune diseases can be exemplified by immunosuppressants;steroids; disease-modifying anti-rheumatic drugs; elastase inhibitors;cannabinoid-2 receptor agonists; prostaglandins; prostaglandin synthaseinhibitors; phosphodiesterase inhibitors; metalloprotease inhibitors;adhesion molecule inhibitors; anti-cytokine protein agents such asanti-TNF-α agents, anti-IL-1 agents, and anti-IL-6 agents; cytokineinhibitors; nonsteroidal antiinflammatories; and anti-CD20 antibodies.

The other drug for supplementing and/or enhancing the prophylacticand/or therapeutic effect of the compounds of the present inventionagainst inflammatory diseases can be exemplified by steroids, elastaseinhibitors, cannabinoid-2 receptor agonists, prostaglandins,prostaglandin synthase inhibitors, phosphodiesterase inhibitors,metalloprotease inhibitors, adhesion molecule inhibitors,anti-leukotrienes, anti-cholinergic agents, thromboxane A2 receptorantagonists, thromboxane synthase inhibitors, xanthine derivatives,expectorants, antibacterials, antihistamines, anti-cytokine proteinagents, cytokine inhibitors, forskolin agents, mediator releaseinhibitors, and nonsteroidal antiinflammatories.

The other drug for supplementing and/or enhancing the prophylacticand/or therapeutic effect of the compounds of the present inventionagainst thromboembolic diseases can be exemplified by thrombolyticagents, heparin, heparinoids, low molecular weight heparins, warfarin,thrombin inhibitors, factor Xa inhibitors, ADP receptor antagonists, andcyclooxygenase inhibitors.

The other drug for supplementing and/or enhancing the prophylacticand/or therapeutic effect of the compounds of the present inventionagainst non-Hodgkin's lymphomas can be exemplified by alkylating agents,antimetabolites, anticancer antibiotics, plant alkaloids, hormones,platinum compounds, anti-CD20 antibodies, and other anticancer agents.

The antihistamines can be exemplified by azelastine hydrochloride,ebastine, epinastine hydrochloride, emedastine fumarate, auranofin,oxatomide, olopatadine hydrochloride, dl-chlorpheniramine maleate,clemastine fumarate, ketotifen fumarate, cimetidine, dimenhydrinate,diphenhydramine hydrochloride, cyproheptadine hydrochloride, cetirizinehydrochloride, desloratadine, terfenadine, famotidine, fexofenadinehydrochloride, bepotastine, bepotastine besilate, mizolastine,mequitazine, mometasone furoate, ranitidine, ranitidine hydrochloride,loratadine, promethazine hydrochloride, and homochlorcyclizinehydrochloride.

The leukotriene antagonists can be exemplified by pranlukast hydrate,montelukast sodium, zafirlukast, ablukast, pobilukast, sulukast,iralukast sodium, verlukast, ritolukast, cinalukast, pirodomast,tomelukast, and doqualast.

The antiallergic drugs can be exemplified by amlexanox, azelastinehydrochloride, israpafant, ibudilast, imitrodast sodium, ebastine,epinastine hydrochloride, emedastine fumarate, oxatomide, ozagrelhydrochloride, olopatadine hydrochloride, cromoglicic acid, sodiumcromoglicate, ketotifen fumarate, seratrodast, cetirizine hydrochloride,suplatast tosilate, tazanolast, terfenadine, domitroban calcium hydrate,tranilast, nedocromil, fexofenadine, fexofenadine hydrochloride,pemirolast potassium, mequitazine, ramatroban, repirinast, andloratadine.

The thromboxane A2 receptor antagonists can be exemplified byseratrodast, domitroban calcium hydrate, and ramatroban.

The thromboxane synthase inhibitors can be exemplified by imitrodastsodium and ozagrel hydrochloride.

The steroids can be exemplified by amcinonide, hydrocortisone sodiumsuccinate, prednisolone sodium succinate, methylprednisolone sodiumsuccinate, ciclesonide, difluprednate, betamethasone propionate,dexamethasone, deflazacort, triamcinolone, triamcinolone acetonide,halcinonide, dexamethasone palmitate, hydrocortisone, flumetasonepivalate, prednisolone butylacetate, budesonide, prasterone sulfate,mometasone furoate, fluocinonide, fluocinolone acetonide,fludroxycortide, flunisolide, prednisolone, alclometasone propionate,clobetasol propionate, dexamethasone propionate, deprodone propionate,fluticasone propionate, beclometasone propionate, betamethasone,methylprednisolone, methylprednisolone suleptanate, methylprednisolonesodium succinate, dexamethasone sodium phosphate, hydrocortisone sodiumphosphate, prednisolone sodium phosphate, diflucortolone valerate,dexamethasone valerate, betamethasone valerate, prednisolone valerateacetate, cortisone acetate, diflorasone acetate, dexamethasone acetate,triamcinolone acetate, paramethason acetate, halopredone acetate,fludrocortisone acetate, prednisolone acetate, methylprednisoloneacetate, clobetasone butyrate, hydrocortisone butyrate, hydrocortisonebutyrate propionate, and betamethasone butyrate propionate.

The immunosuppressants can be exemplified by azathioprine, ascomycin,everolimus, salazosulfapyridine, cyclosporine, cyclophosphamide,sirolimus, tacrolimus, bucillamine, methotrexate, and leflunomide.

The disease-modifying anti-rheumatic drugs can be exemplified byD-penicillamine, actarit, auranofin, salazosulfapyridine,hydroxychloroquine, bucillamine, methotrexate, leflunomide, lobenzaritsodium, aurothioglucose, and sodium aurothiomalate.

The elastase inhibitors can be exemplified by ONO-5046, ONO-6818,MR-889, PBI-1101, EPI-HNE-4, R-665, ZD-0892, ZD-8321, GW-311616,DMP-777, L-659286, L-680833, L-683845, and AE-3763.

The prostaglandins (abbreviated below as “PG”) can be exemplified byPGE1 drugs (examples: alprostadil alfadex, alprostadil), PGI2 drugs(example: beraprost sodium), PG receptor agonists, and PG receptorantagonists. The PG receptor can be exemplified by PGE receptors (EP1,EP2, EP3, and EP4), PGD receptors (DP, CRTH2), PGF receptors (FP), PGI2receptors (IP), and TX receptors (TP).

The prostaglandin synthase inhibitors can be exemplified bysalazosulfapyridine, mesalazine, olsalazine, 4-aminosalicylic acid,JTE-522, auranofin, carprofen, diphenpyramide, flunoxaprofen,flurbiprofen, indometacin, ketoprofen, lornoxicam, loxoprofen,meloxicam, oxaprozin, parsalmide, piproxen, piroxicam, piroxicamcinnamate, zaltoprofen, and pranoprofen.

The phosphodiesterase inhibitors can be exemplified by rolipram,cilomilast, Bay19-8004, NIK-616, roflumilast (BY-217), cipamfylline(BRL-61063), atizoram (CP-80633), ONO-6126, SCH-351591, YM-976,V-11294A, PD-168787, D-4396, and IC-485.

The adhesion molecule inhibitors can be exemplified by α4 integrinantagonist.

The anti-TNF-α agents can be exemplified by anti-TNF-α antibodies,soluble TNF-α receptor, anti-TNF-α receptor antibodies, and solubleTNF-α binding protein and particularly by infliximab and etanercept.

The anti-IL-1 agents can be exemplified by anti-IL-1 antibodies, solubleIL-1 receptor, anti-IL-1Ra antibodies and/or anti-IL-1 receptorantibodies and particularly by anakinra.

The anti-IL-6 agents can be exemplified by anti-IL-6 antibodies, solubleIL-6 receptor, and anti-IL-6 receptor antibodies and particularly bytocilizumab.

The cytokine inhibitors can be exemplified by suplatast tosylate, T-614,SR-31747, and sonatimod.

The anti-cholinergic agents can be exemplified by trihexyphenidyl,trihexyphenidyl hydrochloride, biperiden, and biperiden hydrochloride.

The xanthine derivatives can be exemplified by aminophylline,theophylline, doxofylline, sipamphylline, and diprophylline.

The expectorants can be exemplified by foeniculated ammonia spirit,sodium bicarbonate, bromhexine hydrochloride, carbocysteine, ambroxolhydrochloride, methylcysteine hydrochloride, acetylcysteine, ethylL-cysteine hydrochloride, and tyloxapol.

The antibacterials can be exemplified by sodium cefuroxime, meropenemtrihydrate, netilmicin sulfate, sisomicin sulfate, ceftibuten, PA-1806,IB-367, tobramycin, PA-1420, doxorubicin, astromicin sulfate, andcefetamet pivoxil hydrochloride.

The mediator release inhibitors can be exemplified by tranilast, sodiumcromoglicate, amlexanox, repirinast, ibudilast, dazanolast, andpemirolast potassium.

The thrombolytic agents can be exemplified by alteplase, urokinase,tisokinase, nasaruplase, nateplase, t-PA, pamiteplase, monteplase,prourokinase, and streptokinase.

The heparinoids can be exemplified by fondaparinux.

The low molecular weight heparins can be exemplified by danaparoidsodium, enoxaparin (sodium), nadroparin calcium, bemiparin (sodium),reviparin (sodium), and tinzaparin (sodium).

The thrombin inhibitor can be exemplified by argatroban, ximelagatran,melagatran, dabigatran, bivalirudin, lepirudin, hirudin, and desirudin.

The ADP receptor antagonists can be exemplified by ticlopidinehydrochloride and clopidogrel sulfate.

The cyclooxygenase inhibitors can be exemplified by aspirin.

The alkylating agents can be exemplified by nitrogen mustard N-oxidehydrochloride, cyclophosphamide, ifosfamide, melphalan, thiotepa,carboquone, busulfan, nimustine hydrochloride, dacarbazine, andranimustine.

The antimetabolites can be exemplified by methotrexate, mercaptopurine,6-mercaptopurine riboside, fluorouracil, tegafur, tegafur uracil,carmofur, doxifluridine, cytarabine, enocitabine, tegafur gimestatotastat potassium, gemcitabine hydrochloride, cytarabine ocfosfate,procarbazine hydrochloride, and hydroxycarbamide.

The anticancer antibiotics can be exemplified by actinomycin D,mitomycin C, daunorubicin hydrochloride, doxorubicin hydrochloride,aclarubicin hydrochloride, neocarzinostatin, pirarubicin hydrochloride,epirubicin (hydrochloride), idarubicin hydrochloride, chromomycin A3,bleomycin (hydrochloride), peplomycin sulfate, therarubicin, andzinostatin stimalamer.

The plant alkaloids can be exemplified by vinblastine sulfate,vincristine sulfate, vindesine sulfate, irinotecan hydrochloride,etoposide, flutamide, vinorelbine tartrate, docetaxel hydrate, andpaclitaxel.

The hormones can be exemplified by estramustine phosphate sodium,mepitiostane, epitiostanol, goserelin acetate, fosfestrol(diethylstilbestrol phosphate), tamoxifen citrate, toremifene citrate,fadrozole hydrochloride hydrate, medroxyprogesterone acetate,bicalutamide, leuprorelin acetate, anastrozole, and exemestane.

The platinum compounds can be exemplified by carboplatin, cisplatin, andnedaplatin.

The anti-CD-20 antibodies can be exemplified by rituximab, ibritumomab,and ocrelizumab.

The other anti-cancer agents can be exemplified by L-asparaginase,octreotide acetate, porfimer sodium, and mitoxantrone acetate.

The combined preparation comprising a combination with a compound of thepresent invention not only includes the combined preparations discoveredto date, but also includes combined preparations that may be discoveredin the future.

The compounds of the present invention are generally administeredsystemically or locally as a pharmaceutically effective component in anoral or parenteral form. The oral formulations can be exemplified byliquids for oral administration (for example, elixirs, syrups,pharmaceutically acceptable water-based formulations, suspensions, andemulsions) and solids for oral administration (for example, tablets(including sublingual tablets and orally disintegrating tablets), pills,capsules (including hard capsules, soft capsules, gelatin capsules, andmicrocapsules), powders, granules, and lozenges). The parenteralformulations can be exemplified by solutions (for example, injectables(for example, subcutaneous injectables, intravenous injectables,intramuscular injectables, intraperitoneal injectables, and dripformulations), eye drops (for example, aqueous eye drops (for example,aqueous eye drops, aqueous eye drop suspensions, viscous eye drops, andsolubilized eye drops) and nonaqueous eye drops (for example, nonaqueouseye drops and nonaqueous eye drop suspensions)), topicals (for example,ointments (for example, ophthalmic ointments)), and ear drops). Theseformulations may be controlled release formulations such as rapidrelease formulations, sustained release formulations, and so forth.These formulations can be produced by known methods, for example, by themethods described in The Japanese Pharmacopoeia.

The liquids for oral administration within the sphere of oralformulations can be produced, for example, by dissolving, suspending, oremulsifying the effective component in a commonly used diluent (forexample, purified water, ethanol, or a mixture thereof). These liquidformulations may also contain, for example, a wetting agent, suspendingagent, emulsifying agent, sweetener, flavorant, fragrance, preservative,buffer, and so forth.

The solids for oral administration within the sphere of oralformulations can be prepared by mixing the effective component with, forexample, a vehicle (for example, lactose, mannitol, glucose,microcrystalline cellulose, and starch), a binder (for example,hydroxypropyl cellulose, polyvinylpyrrolidone, and magnesiummetasilicate aluminate), a disintegrant (for example, cellulose calciumglycolate), a lubricant (for example, magnesium stearate), a stabilizer,a dissolution adjuvant (for example, glutamic acid and aspartic acid),and so forth, and formulating according to standard methods. Asnecessary, coating may be carried out with a coating agent (for example,sugar, gelatin, hydroxypropyl cellulose, and hydroxypropyl methylcellulose phthalate) and two or more layers may be applied.

Within the sphere of parenteral formulations, a topical may be producedusing a known method or a formulation in common use. For example, anointment may be prepared by incorporating or melting the effectivecomponent into a base. The ointment base is selected from known ointmentbases or an ointment base in common use. For example, a single selectionfrom the following or a mixture of two or more selections from thefollowing may be used: higher fatty acids and higher fatty acid esters(for example, adipic acid, myristic acid, palmitic acid, stearic acid,oleic acid, adipate esters, myristate esters, palmitate esters, stearateesters, and oleate esters), waxes (for example, beeswax, spermaceti, andceresin), surfactants (for example, polyoxyethylene alkyl etherphosphate esters), higher alcohols (for example, cetanol, stearylalcohol, and cetostearyl alcohol), silicone oils (for example,dimethylpolysiloxane), hydrocarbons (for example, hydrophilicpetrolatum, white petrolatum, purified lanolin, and liquid paraffin),glycols (for example, ethylene glycol, diethylene glycol, propyleneglycol, polyethylene glycol, and macrogol), plant oils (for example,castor oil, olive oil, sesame oil, and turpentine oil), animal oils (forexample, mink oil, egg yolk oil, squalane, and squalene), water,absorption promoters, and anti-irritants. A humectant, preservative,stabilizer, antioxidant, fragrance, and so forth may also beincorporated.

Injectables within the sphere of parenteral formulations encompasssolutions, suspensions, and emulsions as well as solid injectables usedby dissolution or suspension in a solvent at the time of use. Forexample, an injectable may be used in which the effective component isdissolved, suspended, or emulsified in a solvent. For example, distilledwater for injection, physiological saline solution, a plant oil,propylene glycol, polyethylene glycol, an alcohol such as ethanol, or acombination of the preceding may be used for the solvent. The injectablemay also contain a stabilizer, a dissolution adjuvant (for example,glutamic acid, aspartic acid, and Polysorbate 80 (registered trademark)), a suspending agent, an emulsifying agent, a soothing agent, abuffer, a preservative, and so forth. The injectable may be sterilizedin the final step or may be manufactured using aseptic processing. Theinjectable may also be manufactured as a sterile solid form, forexample, a freeze-dried product, and may be used after dissolution indistilled water for injection or another solvent, which is eithersterile or sterilized prior to use.

The dose when a compound of the present invention is used as aneffective component in a drug may be selected as appropriate dependingupon, for example, the symptoms, age, type of formulation, and so forth.In the case of an oral formulation, preferably from 1 mg to 100 mg andmore preferably from 5 mg to 30 mg may be administered from one toseveral times per day (for example, one to three times). In the case ofeye drops, from one to several drops per administration of an ophthalmicsolution having a concentration of preferably 0.000001% to 5% (w/v) andmore preferably 0.00001% to 0.05% (w/v) may be instilled from one toseveral times per day (for example, from one to eight times). In thecase of an ophthalmic ointment, an ophthalmic ointment with aconcentration of preferably 0.000001% to 5% (w/w) and more preferably0.00001% to 0.05% (w/w) may be applied from one to several times per day(for example, one to four times).

Of course, as noted above, the dose will depend upon various conditionsand as a result cases will occur in which an amount less than theaforementioned dosage levels will be sufficient or in which these rangesmust be exceeded.

EXAMPLES

The present invention is described below using examples, but the presentinvention is not limited by these examples.

The solvents given in parentheses for TLC and in the sections onchromatographic separation indicate the elution solvents or developmentsolvents used, and the proportions are volume ratios.

Unless specifically indicated otherwise, the NMR data is data for¹H-NMR.

The solvent used in the measurement is given in parentheses in the NMRsection.

The compound names used in this Description are generally namesgenerated based on IUPAC naming rules or generated using ACD/Name(registered trademark), a computer program from Advanced ChemistryDevelopment, Inc., that performs naming based on IUPAC rules.

Example 1 N,N-dibenzyl-6-chloro-5-nitropyrimidine-4-amine

Dibenzylamine (10.2 g) in a dichloromethane (30 mL) solution was addeddropwise on an ice bath to a dichloromethane (70 mL) solution of4,6-dichloro-5-nitropyrimidine (10 g). This was followed by the additionof triethylamine (14.4 mL) and stirring for 1 hour. Water was then addedto the reaction mixture and the organic layer was subsequently washedwith saturated aqueous sodium chloride solution and dried over anhydroussodium sulfate. The solvent was concentrated under reduced pressure toobtain the title compound (19.2 g) having the physical property valueindicated below.

TLC: Rf 0.50 (hexane:ethyl acetate=7:1).

Example 2 tert-butyl3-{[6-(dibenzylamino)-5-nitropyrimidin-4-yl]amino}azetidine-1-carboxylate

The compound (10.3 g) prepared in Example 1 and tert-butyl3-aminoazetidine-1-carboxylate (5.0 g) were dissolved in dioxane (58mL); triethylamine (8.1 mL) was added; and stirring was then carried outfor 5 hours at 50° C. The reaction mixture was returned to roomtemperature; the solvent was then distilled off; water was added; andextraction was performed with ethyl acetate. The organic layer waswashed with saturated aqueous sodium chloride solution and subsequentlydried over anhydrous sodium sulfate and the solvent was then distilledoff. The residue was purified by silica gel column chromatography toobtain the title compound (10.8 g) having the physical property valueindicated below.

TLC: Rf 0.40 (hexane:ethyl acetate=4:1).

Example 3 tert-butyl3-{[5-amino-6-(dibenzylamino)pyrimidin-4-yl]amino}azetidine-1-carboxylate

An ethyl acetate (360 mL) solution of the compound (17.5 g) prepared inExample 2 was added dropwise to a mixture of zinc (23.3 g) and a 3.0 Maqueous ammonium chloride solution (11.4 g) on an ice bath and thetemperature was immediately raised to room temperature. After stirringfor 2 hours, the reaction mixture was filtered on Celite (trade name)and the solvent was then distilled off. The residue was purified bysilica gel column chromatography to obtain the title compound (12.4 g)having the physical property value indicated below.

TLC: Rf 0.69 (hexane:ethyl acetate=1:1).

Example 4 tert-butyl3-[6-(dibenzylamino)-8-oxo-7,8-dihydro-9H-purin-9-yl]azetidine-1-carboxylate

The compound (8.4 g) prepared in Example 3 and 1,1′-carbonyldiimidazole(5.9 g) were dissolved in tetrahydrofuran (120 mL) followed by stirringfor 15 hours at 60° C. After the solvent had been distilled from thereaction mixture, water was added and extraction with ethyl acetate wasperformed. The organic layer was washed with saturated aqueous sodiumchloride solution and dried over anhydrous sodium sulfate and thesolvent was then distilled off. The residue was purified by silica gelcolumn chromatography to obtain the title compound (7.8 g) having thephysical property value indicated below.

TLC: Rf 0.28 (hexane:ethyl acetate=2:1).

Example 5 tert-butyl3-(6-amino-8-oxo-7,8-dihydro-9H-purin-9-yl)azetidine-1-carboxylate

The compound (7.8 g) prepared in Example 4 was dissolved in methanol(240 mL) and ethyl acetate (50 mL); 20% Pearlman's catalyst (Pd(OH)₂/C)(8.0 g, 100 wt %) was added; replacement with hydrogen was carried out;and stirring was performed for 7.5 hours at 60° C. The reaction mixturewas filtered on Celite (trade name) and the solvent was distilled off toobtain the title compound (5.0 g) having the physical property valueindicated below.

TLC: Rf 0.50 (ethyl acetate).

Example 6 tert-butyl3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]azetidine-1-carboxylate

p-phenoxyphenylboric acid (2.1 g), copper(II) acetate (1.48 g),molecular sieve 4A (2.5 g), and pyridine (0.82 mL) were added at roomtemperature to a dichloromethane (200 mL) suspension of the compound(2.5 g) prepared in Example 5, followed by stirring for 21 hours. Thereaction solution was filtered on Celite (trade name) and the residuewas purified by silica gel column chromatography to obtain the titlecompound (1.3 g) having the physical property value indicated below.

TLC: Rf 0.18 (hexane:ethyl acetate=1:1).

Example 76-amino-9-azetidin-3-yl-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-onedihydrochloride

4 N hydrochloric acid/dioxane (13 mL) was added at room temperature to asuspension in methanol (13 mL) of the compound (1.3 g, 2.76 mmol, 1.0equivalent) prepared in Example 6 and stirring was carried out for 1hour. The solvent was then distilled off to obtain the title compound(1.5 g) having the physical property value indicated below.

TLC: Rf 0.50 (dichloromethane:methanol:28% aqueous ammonia=9:1:0.1).

Example 89-(1-acryloyl-3-azetidinyl)-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

Triethylamine (1.1 mL) was added to a suspension in dichloromethane (16mL) of the compound (1.5 g) prepared in Example 7 and a dichloromethane(10 mL) solution of acryloyl chloride (0.32 mL) was then added dropwiseat −10° C. The solvent was distilled from the reaction mixture followedby the addition of water and extraction with methylene chloride. Theorganic layer was washed with a saturated aqueous sodium chloridesolution and dried over anhydrous sodium sulfate and the solvent wasdistilled off. The residue was purified by silica gel columnchromatography to obtain the title compound (0.8 g) having the physicalproperty values indicated below.

TLC: Rf 0.43 (ethyl acetate);

¹H-NMR (CDCl₃): δ 4.47-4.61, 4.83-4.88, 5.06-5.11, 5.37-5.47, 5.70-5.74,6.21-6.30, 6.36-6.43, 7.07-7.23, 7.35-7.44, 8.24.

Example 8(1) to Example 8(22)

The exemplary compounds given below were obtained by following the sameprocess template as in Example 1→Example 2→Example 3→Example 4→Example5→Example 6→Example 7→Example 8, using 4,6-dichloro-5-nitropyrimidine,using tert-butyl 3-aminoazetidine-1-carboxylate or the correspondingamine derivative in its place, using acryloyl chloride or thecorresponding acid chloride in its place, and using p-phenoxyphenylboricacid or the corresponding boric acid in its place.

Example 8(1)9-[(3R)-1-acryloyl-3-piperidinyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.33 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.64-1.78, 1.90-2.12, 2.55-2.80, 3.10-3.22, 3.63-3.74,3.99-4.12, 4.40-4.55, 4.73-4.82, 5.66-5.72, 6.26-6.31, 6.53-6.65,7.07-7.22, 7.36-7.44, 8.23.

Example 8(2)9-(1-acryloyl-3-azetidinyl)-6-amino-7-[4-(3-chlorophenoxy)-3-methoxyphenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.25 (ethyl acetate);

¹H-NMR (CDCl₃): δ 3.88, 4.47-4.62, 4.83-4.89, 5.07-5.12, 5.39-5.48,5.70-5.74, 6.21-6.30, 6.37-6.43, 6.87-6.91, 6.95-7.00, 7.08-7.13,7.24-7.29, 8.27.

Example 8(3)9-(1-acryloyl-4-piperidinyl)-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.38 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.88-1.96, 2.59-2.80, 3.15-3.27, 4.13-4.22, 4.48,4.58-4.69, 4.85-4.95, 5.69-5.73, 6.27-6.34, 6.59-6.68, 7.07-7.15,7.17-7.22, 7.34-7.44, 8.21.

Example 8(4)9-[(3R)-1-acryloyl-3-pyrrolidinyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.60 (chloroform:methanol:aqueous ammonia=80:10:1);

¹H-NMR (CDCl₃): δ 2.21-2.46, 2.82-3.08, 3.53-3.76, 3.89-4.36, 4.40-4.59,5.11-5.32, 5.64-5.77, 6.34-6.58, 7.04-7.24, 7.34-7.49, 8.18-8.26.

Example 8(5)9-[(3S)-1-acryloyl-3-piperidinyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.71 (chloroform:methanol:aqueous ammonia=80:10:1);

¹H-NMR (CDCl₃): δ 1.52-1.80, 1.88-2.17, 2.51-2.84, 3.07-3.26, 3.59-3.76,3.94-4.16, 4.38-4.63, 4.66-4.90, 5.61-5.80, 6.21-6.40, 6.48-6.72,7.03-7.25, 7.33-7.50, 8.23.

Example 8(6)9-[(3S)-1-acryloyl-3-pyrrolidinyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.50 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 2.23-2.43, 2.84-3.04, 3.55-3.73, 3.91-4.08, 4.12-4.19,4.26-4.32, 4.47-4.49, 5.13-5.27, 5.66-5.73, 6.35-6.54, 7.03-7.28,7.33-7.43, 8.20-8.22.

Example 8(7)9-(1-acryloyl-3-azetidinyl)-6-amino-7-[4-(3-chlorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.53 (chloroform:methanol:aqueous ammonia=9:1:0.01);

¹H-NMR (CDCl₃): δ 4.46-4.61, 4.85, 5.08, 5.42, 5.71, 6.24, 6.38, 6.96,7.08, 7.13-7.18, 7.32, 7.42, 8.24.

Example 8(8)9-(1-acryloyl-3-azetidinyl)-6-amino-7-(3-methoxy-4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.18 (ethyl acetate);

¹H-NMR (CDCl₃): δ 3.90, 4.45-4.65, 4.80-4.90, 5.05-5.15, 5.35-5.45,5.72, 6.26, 6.39, 6.90-7.40, 8.25.

Example 8(9)9-(1-acryloyl-3-azetidinyl)-6-amino-7-(3-fluoro-4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.27 (ethyl acetate);

¹H-NMR (CDCl₃): δ 4.45-4.65, 4.80-4.90, 5.05-5.15, 5.35-5.45, 5.72,6.25, 6.40, 7.05-7.42, 8.26.

Example 8(10)9-(1-acryloyl-3-azetidinyl)-6-amino-7-[4-(3,4-dichlorophenoxy)-3-methoxyphenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.20 (ethyl acetate);

¹H-NMR (CD₃OD): δ 3.82, 4.47, 4.59, 4.72, 4.77-4.95, 5.00-5.09,5.40-5.53, 5.77, 6.28, 6.41, 6.91, 7.09, 7.28, 7.43, 8.17.

Example 8(11)9-(1-acryloyl-3-azetidinyl)-6-amino-7-[4-(3-fluorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.73 (ethyl acetate:methanol:triethylamine=9:1:0.1);

¹H-NMR (CDCl₃): δ 4.50, 4.59, 4.85, 5.08, 5.42, 5.72, 6.25, 6.40,6.78-6.93, 7.17, 7.33, 7.43, 8.26.

Example 8(12)9-(1-acryloyl-3-azetidinyl)-6-amino-7-[4-(3,5-difluorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.78 (ethyl acetate:methanol:triethylamine=9:1:0.1);

¹H-NMR (CDCl₃): δ 4.50, 4.59, 4.85, 5.08, 5.42, 5.71, 6.25, 6.39,6.57-6.62, 7.19, 7.45, 8.26.

Example 8(13)9-(1-acryloyl-3-azetidinyl)-6-amino-7-(2-fluoro-4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.47 (methanol:ethyl acetate=1:19);

¹H-NMR (CDCl₃): δ 4.40-4.60, 4.80-4.90, 5.00-5.10, 5.35-5.45, 5.72,6.25, 6.39, 6.80-7.50, 8.24.

Example 8(14)9-[(1-acryloyl-4-piperidinyl)methyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.50 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.27-1.44, 1.72-1.86, 2.18-2.37, 2.66, 3.04, 3.90,4.00, 4.51, 4.67, 5.66, 6.24, 6.56, 7.03-7.24, 7.32-7.46, 8.25.

Example 8(15)6-amino-9-[1-(chloroacetyl)-3-azetidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.41 (ethyl acetate:methanol=19:1); 1H-NMR (CDCl₃): δ 3.96-4.05,4.44-4.53, 4.62-4.72, 4.80-4.87, 5.08-5.16, 5.38-5.48, 7.05-7.24,7.35-7.43, 8.24.

Example 8(16)6-amino-9-[(3R)-1-(chloroacetyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.49 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 2.20-2.48, 2.78-3.01, 3.50-3.63, 3.64-3.81, 3.90-4.19,4.26-4.36, 4.51, 5.14-5.32, 7.02-7.26, 7.27-7.46, 8.21.

Example 8(17)6-amino-9-{(3R)-1-[(2E)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.41 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.88, 2.19-2.43, 2.82-3.05, 3.50-3.74, 3.88-4.18,4.20-4.30, 4.48, 5.08-5.16, 6.13, 6.95, 7.05-7.23, 7.34-7.46, 8.22.

Example 8(18)6-amino-9-{[1-(chloroacetyl)-4-piperidinyl]methyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.54 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.14-1.59, 1.70-1.87, 2.19-2.37, 2.65, 3.10,3.81-3.95, 3.91, 4.06, 4.50, 4.51-4.63, 7.05-7.23, 7.34-7.46, 8.25.

Example 8(19)9-[(l-acryloyl-4-methyl-4-piperidinyl)methyl]-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.55 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.15, 1.37-1.85, 3.21-3.50, 3.70-3.99, 4.03-4.23,4.52-4.73, 5.64, 6.24, 6.56, 6.97-7.23, 7.31-7.50, 8.21.

Example 8(20)6-amino-9-{1-[(2E)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.63 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.89, 4.46, 4.51, 4.79, 5.03, 5.20, 5.39, 5.94, 6.93,7.07-7.10, 7.19, 7.36-7.43, 8.23.

Example 8(21)9-{[(3R)-1-acryloyl-3-pyrrolidinyl]methyl}-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-8-one

TLC: Rf 0.49 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 1.77-1.97, 2.02-2.19, 2.83-3.01, 3.38-3.62, 3.66-3.83,3.98-4.11, 4.42-4.54, 5.61-5.70, 6.30-6.45, 7.04-7.23, 7.36-7.43, 8.23.

Example 8(22)9-{[(2S)-1-acryloyl-2-pyrrolidinyl]methyl}-6-amino-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.44 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 1.87-2.28, 3.46-3.78, 3.86-3.96, 3.99-4.10, 4.15-4.23,4.40, 4.45-4.60, 4.81-4.91, 5.54-5.72, 6.16-6.44, 6.86-6.97, 7.04-7.22,7.36-7.45, 8.21-8.26.

Example 9 tert-butyl(3R)-3-{[6-(dibenzylamino)-5-nitropyrimidin-4-yl]amino}piperidine-1-carboxylate

The title compound (1.44 g) having the physical property value givenbelow was obtained by following the same process template as in Example2 using the compound (1.5 g) produced in Example 1 and using tert-butyl(3R)-3-aminopiperidine-1-carboxylate (0.85 g) in place of the tert-butyl3-aminoazetidine-1-carboxylate.

TLC: Rf 0.23 (hexane:ethyl acetate=9:1).

Example 106-amino-7-(4-phenoxyphenyl)-9-[(3R)-piperidin-3-yl]-7,9-dihydro-8H-purin-8-onedihydrochloride

The title compound (155 mg) having the physical property value givenbelow was obtained by following the same process template as in Example3→Example 4→Example 5→Example 6→Example 7, using the compound producedin Example 9 and using p-phenoxyphenylboric acid (154 mg).

TLC: Rf 0.68 (methanol:dichloromethane:aqueous ammonia=80:20:4).

Example 116-amino-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-piperidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

Triethylamine (1.40 mL) was added to a dichloromethane (6 mL) solutionof the compound (1.20 g) produced in Example 10. 4-bromocrotonylchloride (0.5 M dichloroethane solution, 5.04 mL) was added on an icebath and the reaction mixture was stirred for 30 minutes on an ice bath.After concentration, a saturated aqueous sodium bicarbonate solution wasadded and ethyl acetate extraction was performed. The obtained organiclayer was washed with a saturated aqueous sodium chloride solution andwas dried over sodium sulfate, filtered, and concentrated.Tetrahydrofuran (6 mL) and dimethylamine (2.0 M tetrahydrofuransolution, 6.3 mL) were added to the residue and heating was carried outfor 2 hours at 50° C. This was followed by filtration and thenconcentration. The resulting residue was purified by silica gel columnchromatography (hexane:ethyl acetate=1:1→ethyl acetate→ethylacetate:methanol:triethylamine=18:2:1) to yield the title compound (0.66g) having the physical property values indicated below.

TLC: Rf 0.40 (ethyl acetate:methanol:triethylamine=18:2:1);

¹H-NMR (CDCl₃): δ 1.52-1.77, 1.87-2.00, 2.00-2.12, 2.18-2.35, 2.53-2.85,2.93-3.22, 3.57-3.74, 3.97-4.17, 4.36-4.60, 4.66-4.89, 6.35-6.55,6.76-6.94, 7.03-7.30, 7.32-7.48, 8.21.

Examples 11(1) to 11(26)

The exemplary compounds given below were obtained by following the sameprocess template as in Example 9→Example 10→Example 11, using thecompound produced in Example 1, using tert-butyl(3R)-3-aminopiperidine-1-carboxylate or the corresponding aminederivative in its place, using p-phenoxyphenylboric acid or thecorresponding boric acid in its place, and using dimethylamine or thecorresponding amine derivative in its place.

Example 11(1)6-amino-9-{l-[(2E)-4-(dimethylamino)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.21 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.27, 3.08-3.10, 4.45-4.60, 4.80-4.86, 5.05-5.10,5.36-5.44, 6.06-6.12, 6.87-6.96, 7.07-7.15, 7.17-7.23, 7.35-7.44, 8.24.

Example 11(2)6-amino-9-{l-[(2E)-4-(4-morpholinyl)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.48, 3.14-3.16, 3.71-3.73, 4.44-4.59, 4.80-4.86,5.04-5.09, 5.36-5.46, 6.08-6.13, 6.86-6.95, 7.07-7.15, 7.17-7.22,7.34-7.42, 8.23.

Example 11(3)6-amino-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.21 (dichloromethane:methanol=17:3);

¹H-NMR (CDCl₃): δ 2.20-2.42, 2.84-3.14, 3.52-3.76, 3.90-4.20, 4.22-4.34,4.42-4.51, 5.10-5.29, 6.20-6.38, 6.86-7.01, 7.03-7.24, 7.37-7.46,8.20-8.23.

Example 11(4)6-amino-7-(4-phenoxyphenyl)-9-{1-[(2E)-4-(1-piperidinyl)-2-butenoyl]-3-azetidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.32-1.47, 1.57-1.64, 2.38-2.44, 3.13-3.14, 4.44-4.60,4.80-4.86, 5.04-5.09, 5.35-5.46, 6.05-6.11, 6.89-6.99, 7.07-7.15,7.17-7.23, 7.35-7.44, 8.24.

Example 11(5)6-amino-7-(4-phenoxyphenyl)-9-{1-[(2E)-4-(4-thiomorpholinyl)-2-butenoyl]-3-azetidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.58 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.66-2.75, 3.15-3.17, 4.44-4.59, 4.80-4.86, 5.04-5.09,5.36-5.44, 6.05-6.11, 6.85-6.94, 7.07-7.15, 7.17-7.23, 7.35-7.44, 8.24.

Example 11(6)6-amino-9-{1-[(2E)-4-(3-oxo-1-piperazinyl)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.16 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.67-2.71, 3.19, 3.22-3.24, 3.36-3.40, 4.45-4.59,4.81-4.87, 5.05-5.10, 5.37-5.47, 6.05, 6.10-6.16, 6.84-6.93, 7.07-7.15,7.17-7.23, 7.36-7.44, 8.24.

Example 11(7)6-amino-7-(4-phenoxyphenyl)-9-{1-[(2E)-4-(1-pyrrolidinyl)-2-butenoyl]-3-azetidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.39 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.79-1.83, 2.50-2.65, 3.28-3.31, 4.44-4.60, 4.80-4.85,5.04-5.09, 5.35-5.45, 6.10-6.15, 6.90-6.99, 7.06-7.14, 7.16-7.22,7.35-7.43, 8.23.

Example 11(8) 6-amino-9-(1-{(2E)-4-[(2-hydroxyethyl)(methyl)amino]-2-butenoyl}-3-azetidinyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.40 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 2.29, 2.57-2.61, 3.23-3.26, 3.59-3.63, 4.44-4.59,4.80-4.86, 5.04-5.09, 5.36-5.46, 6.06-6.13, 6.86-6.95, 7.07-7.15,7.16-7.26, 7.35-7.42, 8.24.

Example 11(9)6-amino-7-(4-phenoxyphenyl)-9-{(3R)-1-[(2E)-4-(1-pyrrolidinyl)-2-butenoyl]-3-pyrrolidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.42 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.74-1.92, 2.15-2.43, 2.54-2.70, 2.82-3.06, 3.32,3.49-3.77, 3.91-4.20, 4.23-4.36, 4.51, 5.10-5.28, 6.36, 6.91-7.04,7.06-7.24, 7.37-7.45, 8.21.

Example 11(10)6-amino-7-(4-phenoxyphenyl)-9-{(3R)-1-[(2E)-4-(1-piperidinyl)-2-butenoyl]-3-pyrrolidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.39 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.35-1.50, 1.51-1.65, 2.20-2.48, 2.82-3.06, 3.12,3.52-3.75, 3.88-4.19, 4.21-4.33, 4.51, 5.09-5.28, 6.27, 6.88-7.04,7.05-7.23, 7.34-7.44, 8.22.

Example 11(11)6-amino-9-{(3R)-1-[(2E)-4-(4-morpholinyl)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.51 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 2.19-2.54, 2.83-3.05, 3.14, 3.51-3.77, 3.89-4.19,4.21-4.32, 4.57, 5.10-5.28, 6.31, 6.87-6.99, 7.04-7.23, 7.34-7.46, 8.21.

Example 11(12)6-amino-7-(4-phenoxyphenyl)-9-{(3R)-1-[(2E)-4-(4-thiomorpholinyl)-2-butenoyl]-3-pyrrolidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.54 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 2.10-2.43, 2.50-2.78, 2.82-3.06, 3.15, 3.51-3.74,3.89-4.19, 4.22-4.33, 4.51, 5.09-5.28, 6.29, 6.86-6.99, 7.04-7.23,7.35-7.47, 8.22.

Example 11(13)6-amino-9-[(3R)-1-{(2E)-4-[ethyl(methyl)amino]-2-butenoyl}-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.34 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.05-1.19, 2.20-2.43, 2.35, 2.59, 2.82-3.08, 3.31,3.50-3.77, 3.90-4.20, 4.23-4.33, 4.50, 5.10-5.28, 6.38, 6.88-6.99,7.04-7.23, 7.35-7.46, 8.21.

Example 11(14)6-amino-9-{(3S)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.38 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 2.20-2.42, 2.83-3.14, 3.52-3.74, 3.92-4.20, 4.24-4.32,4.42-4.51, 5.13-5.25, 6.20-6.38, 6.87-7.00, 7.05-7.24, 7.35-7.43,8.20-8.23.

Example 11(15)6-amino-9-{1-[(2E)-4-(diethylamino)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.47 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.01-1.06, 2.51-2.58, 3.25-3.27, 4.44-4.59, 4.81-4.86,5.04-5.09, 5.36-5.46, 6.07-6.13, 6.91-7.00, 7.07-7.15, 7.17-7.23,7.36-7.44, 8.24.

Example 11(16)6-amino-9-(1-{(2E)-4-[ethyl(methyl)amino]-2-butenoyl}-3-azetidinyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.50 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.05-1.09, 2.24, 2.41-2.48, 3.15-3.18, 4.44-4.59,4.80-4.86, 5.04-5.09, 5.36-5.46, 6.05-6.12, 6.89-6.98, 7.07-7.15,7.17-7.23, 7.36-7.44, 8.24.

Example 11(17)2-{[(2E)-4-{3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]-1-azetidinyl}-4-oxo-2-buten-1-yl](methyl)amino}acetamide

TLC: Rf 0.30 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.35, 3.04, 3.23-3.25, 4.45-4.60, 4.81-4.87,5.04-5.09, 5.37-5.47, 5.61, 6.05-6.11, 6.84-6.94, 6.99, 7.07-7.15,7.17-7.23, 7.35-7.44, 8.24.

Example 11(18)6-amino-9-{(3R)-1-[(2E)-4-(diethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.52 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 0.96-1.11, 2.21-2.43, 2.45-2.63, 2.83-3.05, 3.29,3.52-3.78, 3.90-4.20, 4.22-4.33, 4.49, 5.10-5.28, 6.33, 6.90-7.04,7.05-7.23, 7.34-7.47, 8.21.

Example 11(19) 6-amino-9-[(3R)-1-{(2E)-4-[(2-hydroxyethyl)(methyl)amino]-2-butenoyl}-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.18 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 2.21-2.62, 2.85-3.08, 3.21-3.28, 3.52-3.74, 3.92-4.17,4.25-4.33, 4.61-4.63, 5.13-5.30, 6.24-6.37, 6.87-6.98, 7.07-7.23,7.35-7.44, 8.19-8.22.

Example 11(20)6-amino-9-{1-[(2E)-4-(4-hydroxy-1-piperidinyl)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.30 (dichloromethane:methanol:triethylamine=9:1:0.1);

¹H-NMR (CDCl₃): δ 1.30-1.75, 1.82-1.97, 2.10-2.24, 2.65-2.82, 3.05-3.20,3.61-3.78, 4.40-4.60, 4.78-4.86, 5.00-5.11, 5.33-5.46, 6.02-6.12,6.84-6.98, 7.05-7.24, 7.35-7.43, 8.23.

Example 11(21)6-amino-7-[4-(3-chlorophenoxy)phenyl]-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.28 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 2.17-2.43, 2.25, 2.83-3.05, 3.08, 3.51-3.76,3.91-4.20, 4.22-4.33, 4.50, 5.10-5.18, 6.29, 6.88-7.01, 7.09, 7.10-7.20,7.33, 7.42, 8.23.

Example 11(22)6-amino-9-[(3S)-1-{(2E)-4-[ethyl(methyl)amino]-2-butenoyl}-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.44 (ethyl acetate:methanol:triethylamine=18:2:1);

¹H-NMR (CDCl₃): δ 1.00-1.14, 2.18-2.32, 2.32-2.53, 2.82-3.06, 3.10-3.23,3.50-3.77, 3.86-4.09, 4.09-4.35, 4.37-4.58, 5.06-5.32, 6.20-6.41,6.87-7.03, 7.03-7.25, 7.31-7.50, 8.14-8.30.

Example 11(23)6-amino-9-{(3S)-1-[(2E)-4-(diethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.44 (ethyl acetate:methanol:triethylamine=18:2:1);

¹H-NMR (CDCl₃): δ 0.97-1.11, 2.19-2.46, 2.47-2.65, 2.85-3.05, 3.21-3.35,3.50-3.76, 3.89-4.09, 4.09-4.33, 4.40-4.55, 5.10-5.28, 6.21-6.43,6.88-7.05, 7.05-7.24, 7.34-7.48, 8.17-8.27.

Example 11(24)6-amino-9-{1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.38 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 2.20-2.42, 2.83-3.14, 3.52-3.74, 3.92-4.20, 4.24-4.32,4.42-4.51, 5.13-5.25, 6.20-6.38, 6.87-7.00, 7.05-7.24, 7.35-7.43,8.20-8.23.

Example 11(25)6-amino-9-({(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}methyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.28 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.75-1.97, 2.00-2.16, 2.20-2.26, 2.83-3.00, 3.02-3.10,3.38-3.62, 3.69-3.82, 3.98-4.08, 4.50-4.60, 6.19-6.31, 6.82-6.96,7.04-7.22, 7.35-7.43, 8.23.

Example 11(26)6-amino-9-({(2S)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-2-pyrrolidinyl}methyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.30 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (CDCl₃): δ 1.86-2.27, 2.98-3.02, 3.03-3.17, 3.45-3.76, 3.88-3.95,3.98-4.08, 4.15-4.23, 4.39-4.42, 4.47-4.57, 4.80-4.91, 6.09-6.19,6.66-6.83, 6.94-7.03, 7.04-7.23, 7.35-7.46, 8.20-8.27.

Example 12 Dimethylaminoacetaldehyde Disulfite

Water (1 mL) and concentrated hydrochloric acid (2.4 mL) were added todimethylaminoacetaldehyde diethyl acetal (2.00 g) and heating wascarried out for 3 hours at 40° C. Sodium disulfite (3.0 M aqueoussolution, 3.6 mL) and ethanol (10 mL) were added on an ice bath andstirring was performed for 1 hour and 30 minutes at room temperature.Filtration was carried out followed by concentration. The resultingresidue was evaporated to dryness under reduced pressure at 60° C. toobtain the title compound (1.29 g).

Example 13 tert-butyl(3R)-3-{[6-(dibenzylamino)-5-nitropyrimidin-4-yl]amino}pyrrolidine-1-carboxylate

The title compound (27.0 g) having the physical property value givenbelow was obtained by following the same process template as in Example2, using the compound (19 g) produced in Example 1 and using tert-butyl(3R)-3-aminopyrrolidine-1-carboxylate (10.5 g) in place of thetert-butyl 3-aminoazetidine-1-carboxylate.

TLC: Rf 0.29 (hexane:ethyl acetate=4:1).

Example 146-amino-7-(4-phenoxyphenyl)-9-[(3R)-pyrrolidin-3-yl]-7,9-dihydro-8H-purin-8-onedihydrochloride

The title compound (945 mg) having the physical property value indicatedbelow was obtained by following the same process template as in Example10, using the compound (9.0 g) produced in Example 13 and usingp-phenoxyphenylboric acid (2.1 g).

TLC: Rf 0.43 (dichloromethane:methanol:aqueous ammonia=8:1:0.1).

Example 15 diethyl(2-{[(3R)-3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]pyrrolidin-1-yl}-2-oxoethyl)phosphonate

1,1′-carbonyldiimidazole (0.827 g) was added to tetrahydrofuran (5 mL)and heating to 40° C. was carried out. To this solution was addeddiethylphosphonoacetic acid (1.00 g) dissolved in tetrahydrofuran (5mL). Stirring was performed for 30 minutes at 40° C. The thusly producedsolution (0.28 mL), the compound (50 mg) produced in Example 14, andtriethylamine (0.03 mL) were added to tetrahydrofuran (0.5 mL) andstirring was performed for 1 day at room temperature. Water was addedand extraction with ethyl acetate was carried out. The obtained organiclayer was washed with a saturated aqueous sodium chloride solution,dried over sodium sulfate, then filtered, and concentrated. Theresulting residue was evaporated to dryness under reduced pressure toobtain the title compound (53 mg) having the physical property valueindicated below.

TLC: Rf 0.46 (chloroform:methanol:28% aqueous ammonia=80:10:1).

Example 166-amino-9-{(3R)-1-[(2Z)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

The compound (53 mg) prepared in Example 12 was added to ethanol (0.9mL) and lithium chloride (4.2 mg) was added. Potassium hydroxide (45%aqueous solution, 0.056 mL) was added on an ice bath and the compound(25 mg) prepared in Example 15 dissolved in water (0.25 mL) was added.Stirring was carried out for 30 minutes on an ice bath. Another additionwas made of the compound (7.5 mg) prepared in Example 15 dissolved inwater (0.25 mL). Stirring was carried out for 2 hours on an ice bath,after which stirring was carried out for 1 day at room temperature.Extraction was performed with ethyl acetate, and the organic layer waswashed with a saturated aqueous sodium chloride solution and wasconcentrated under reduced pressure. The resulting residue was purifiedby thin-layer chromatography (dichloromethane:methanol:28% aqueousammonium=80:10:1) to obtain the title compound (7.7 mg) having thephysical property values indicated below.

TLC: Rf 0.43 (chloroform:methanol:aqueous ammonia=80:10:1);

¹H-NMR (CDCl₃): δ 2.14-2.45, 2.81-3.03, 3.43-3.68, 3.80-4.06, 4.06-4.28,4.42-4.62, 5.11-5.29, 5.97-6.24, 7.03-7.25, 8.17-8.28.

Example 17 tert-butyl4-({[6-(dibenzylamino)-5-nitropyrimidin-4-yl]amino}methyl)piperidine-1-carboxylate

The title compound (68.3 g) having the physical property value indicatedbelow was obtained by following the same process template as in Example2, using the compound (45.5 g) produced in Example 1 and usingtert-butyl 4-(aminomethyl)piperidine-1-carboxylate (27.5 g) in place ofthe tert-butyl 3-aminoazetidine-1-carboxylate.

TLC: Rf 0.56 (hexane:ethyl acetate=2:1).

Example 186-amino-7-(4-phenoxyphenyl)-9-(piperidin-4-ylmethyl)-7,9-dihydro-8H-purin-8-onedihydrochloride

The title compound (1.66 g) having the physical property value indicatedbelow was obtained by following the same process template as in Example10, using the compound produced in Example 17 and usingp-phenoxyphenylboric acid (19 g).

TLC: Rf 0.10 (ethyl acetate:methanol:triethylamine=18:2:1).

Example 196-amino-9-{[1-(2-butynoyl)-4-piperidinyl]methyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

2-butynoic acid (34 mg), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimidehydrochloride (EDC) (78 mg), 1-hydroxybenzotriazole (HOBt) (62 mg), andtriethylamine (114 μL) were added to a dimethylformamide (3 mL) solutionof the compound (100 mg) prepared in Example 18, followed by stirringfor 3 hours at room temperature. Water was added to the reaction mixtureand extraction with ethyl acetate was performed. The organic layer waswashed with a saturated aqueous sodium bicarbonate solution and with asaturated aqueous sodium chloride solution and was then dried overanhydrous sodium sulfate and the solvent was distilled off. The residuewas purified by thin-layer chromatography (dichloromethane:methanol:28%aqueous ammonia=90:10:1) to obtain the title compound (75 mg) having thephysical property values indicated below.

TLC: Rf 0.43 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.21-1.45, 1.71-1.83, 1.99, 2.18-2.36, 2.59-2.72,2.99-3.94, 4.34-4.61, 7.05-7.24, 7.36-7.43, 8.24.

Examples 19(1) to 19(49)

The exemplary compounds given below were obtained by following the sameprocess template as in Example 9→Example 10→Example 19, using thecompound prepared in Example 1, using tert-butyl(3R)-3-aminopiperidine-1-carboxylate or the corresponding aminederivative in its place, and using p-phenoxyphenylboric acid or thecorresponding boric acid in its place.

Example 19(1)6-amino-9-[1-(2-butynoyl)-3-azetidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.38 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.99, 4.39-4.57, 4.75-4.83, 4.97-5.05, 5.32-5.43,7.05-7.24, 7.35-7.43, 8.24.

Example 19(2)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.68 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.23-2.39, 2.80-3.01, 3.50-3.63, 3.67-3.80,3.86-4.02, 4.03-4.18, 4.23-4.33, 4.42-4.51, 5.11-5.25, 7.04-7.23,7.34-7.45, 8.20-8.23.

Example 19(3)6-amino-9-[(3S)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.27 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.89-4.00, 4.03-4.18, 4.23-4.34, 4.42-4.51, 5.13-5.25, 7.05-7.24,7.35-7.43, 8.20-8.23.

Example 19(4)6-amino-7-(4-phenoxyphenyl)-9-[(3R)-1-propioloyl-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.51 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 2.25-2.43, 2.82-3.01, 3.05, 3.52-3.66, 3.73-3.85,3.92-4.04, 4.07-4.23, 4.31-4.40, 4.48, 5.17-5.27, 7.06-7.23, 7.34-7.47,8.23.

Example 19(5)(2E)-4-{3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]-1-azetidinyl}-4-oxo-N-phenyl-2-butenamide

TLC: Rf 0.56 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 4.43-4.72, 4.87-4.96, 5.10-5.20, 5.40-5.52, 6.20,6.36, 7.04-7.43, 7.72, 8.22, 12.49.

Example 19(6)(2E)-4-{3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]-1-azetidinyl}-4-oxo-2-butenamide

TLC: Rf 0.50 (ethyl acetate:methanol:triethylamine=9:1:0.5);

¹H-NMR (DMSO): δ 4.29-4.35, 4.57-4.70, 4.86-4.91, 5.28-5.37, 5.82,6.77-6.88, 7.10-7.20, 7.39-7.46, 7.86, 8.14.

Example 19(7)6-amino-7-(4-phenoxyphenyl)-9-[(1-propioloyl-4-piperidinyl)methyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.38 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.26-1.50, 1.74-1.88, 2.19-2.38, 2.69, 3.09, 3.10,3.91, 4.41, 4.49, 4.56, 7.05-7.23, 7.35-7.44, 8.25.

Example 19(8)(2E)-4-{(3R)-3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]-1-pyrrolidinyl}-4-oxo-2-butenamide

TLC: Rf 0.41 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 2.23-2.47, 2.81-3.02, 3.45-3.82, 3.99-4.23, 4.35-4.53,5.16-5.31, 5.50-5.65, 5.84-6.00, 6.97-7.30, 7.34-7.43, 8.19-8.22.

Example 19(9)6-amino-9-[1-(2-butynoyl)-3-pyrrolidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.27 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.89-4.00, 4.03-4.18, 4.23-4.34, 4.42-4.51, 5.13-5.25, 7.05-7.24,7.35-7.43, 8.20-8.23.

Example 19(10) ethyl(2E)-4-{3-[6-amino-8-oxo-7-(4-phenoxyphenyl)-7,8-dihydro-9H-purin-9-yl]-1-azetidinyl}-4-oxo-2-butenoate

TLC: Rf 0.35 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.30-1.35, 4.22-4.30, 4.48-4.68, 4.84-4.90, 5.12-5.17,5.40-5.50, 6.86-7.06, 7.07-7.16, 7.18-7.23, 7.35-7.44, 8.24.

Example 19(11)6-amino-9-{1-[(2E)-4-phenoxy-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.55 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 4.38-4.60, 4.68-4.79, 4.80-4.89, 4.99-5.11, 5.32-5.46,6.24-6.34, 6.87-7.43, 8.22.

Example 19(12)6-amino-9-{1-[(2E)-4-hydroxy-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.21 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.82-1.90, 4.35-4.41, 4.43-4.62, 4.79-4.87, 5.03-5.12,5.36-5.47, 6.19-6.26, 6.98-7.23, 7.35-7.44, 8.23.

Example 19(13)6-amino-9-{1-[(2E)-2,4-pentadienoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.47 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 4.42-4.67, 4.80-4.90, 5.02-5.13, 5.33-5.50, 5.61,6.02, 6.39-6.55, 7.04-7.27, 7.32-7.48, 8.24.

Example 19(14)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(cyclohexyloxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.44 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.22-2.46, 1.48-1.70, 1.72-1.88, 1.94-2.05, 2.22-2.40,2.80-3.01, 3.51-3.62, 3.68-3.80, 3.87-4.00, 4.02-4.18, 4.22-4.37,4.42-4.55, 5.11-5.24, 6.98-7.05, 7.29-7.36, 8.19-8.23.

Example 19(15)6-amino-9-({(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]-3-pyrrolidinyl}methyl)-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.28 (chloroform:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.75-1.97, 2.00-2.16, 2.20-2.26, 2.83-3.00, 3.02-3.10,3.38-3.62, 3.69-3.82, 3.98-4.08, 4.50-4.60, 6.19-6.31, 6.82-6.96,7.04-7.22, 7.35-7.43, 8.23.

Example 19(16)6-amino-9-{[(2S)-1-(2-butenoyl)-2-pyrrolidinyl]methyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.75, 3.80-2.27, 3.58-3.73, 3.94-4.04, 4.06-4.19,4.39-4.55, 4.66-4.82, 7.03-7.22, 7.35-7.46, 8.22-8.24.

Example 19(17)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-chlorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.45 (dichloromethane:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.51, 5.13-5.25, 6.93-7.00,7.05-7.09, 7.10-7.20, 7.26-7.35, 7.38-7.44, 8.20-8.24.

Example 19(18)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-fluorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.52 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.51-3.62, 3.68-3.80,3.88-4.00, 4.03-4.19, 4.23-4.34, 4.40-4.55, 5.13-5.25, 6.77-6.93,7.12-7.20, 7.28-7.44, 8.20-8.25.

Example 19(19)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3,5-difluorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.52-3.63, 3.68-3.80,3.88-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.54, 5.13-5.25, 6.53-6.66,7.17-7.27, 7.41-7.48, 8.21-8.26.

Example 19(20)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(3-fluoro-4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.41 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.00, 3.52-3.63, 3.68-3.80,3.88-4.00, 4.03-4.19, 4.23-4.34, 4.46-4.57, 5.13-5.25, 7.02-7.21,7.29-7.42, 8.21-8.26.

Example 19(21)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-fluorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.26 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.54, 5.12-5.25, 6.99-7.16,7.35-7.41, 8.20-8.24.

Example 19(22)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-chlorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.28 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.02, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.54, 5.12-5.25, 7.00-7.06,7.09-7.17, 7.30-7.43, 8.20-8.24.

Example 19(23)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.35 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.51, 5.13-5.25, 6.84-6.92,6.99-7.03, 7.07-7.15, 7.24-7.31, 7.36-7.41, 8.20-8.24.

Example 19(24)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.33 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.22-2.42, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.55, 5.13-5.25, 6.95-7.02,7.04-7.16, 7.17-7.23, 7.33-7.41, 8.20-8.24.

Example 19(25)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-{4-[3-(trifluoromethyl)phenoxy]phenyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.33 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.22-2.42, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.42-4.55, 5.13-5.25, 7.12-7.20,7.22-7.28, 7.35-7.38, 7.40-7.57, 8.20-8.24.

Example 19(26)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-methoxyphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.23 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.83, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.41-4.55, 5.13-5.25, 6.89-6.97,7.00-7.08, 7.30-7.38, 8.19-8.23.

Example 19(27)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-methoxyphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.28 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.47-3.62, 3.69-3.83,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.41-4.55, 5.13-5.25, 6.62-6.69,6.71-6.78, 7.10-7.19, 7.24-7.35, 7.36-7.42, 6.89-6.97, 7.00-7.08,7.30-7.38, 8.20-8.24.

Example 19(28)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-isopropylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.31 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.20-1.34, 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.47-3.63,3.69-3.79, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.46-4.57, 5.13-5.25,6.98-7.03, 7.06-7.15, 7.21-7.30, 7.33-7.41, 8.20-8.24.

Example 19(29)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-isopropylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.40 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.20-1.28, 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.47-3.63,3.69-3.80, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.46-4.57, 5.13-5.25,6.84-6.90, 6.98, 7.04-7.16, 7.27-7.41, 8.20-8.24.

Example 19(30)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(2-isopropylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.37 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.20-1.27, 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.16-3.30,3.47-3.63, 3.69-3.80, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.46-4.56,5.13-5.25, 6.93-7.00, 7.01-7.08, 7.17-7.24, 7.32-7.41, 8.20-8.24.

Example 19(31)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(2-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.64 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24, 2.24-2.40, 2.81-3.01, 3.47-3.63,3.69-3.80, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.44-4.55, 5.13-5.25,6.98-7.05, 7.10-7.18, 7.20-7.40, 8.20-8.24.

Example 19(32)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3-chlorophenoxy)-3-methoxyphenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.33 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.82-3.01, 3.47-3.63, 3.69-3.81,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.48-4.60, 5.14-5.26, 6.85-6.92,6.95-7.01, 7.05-7.14, 7.22-7.29, 8.23-8.27.

Example 19(33)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3,4-dichlorophenoxy)-3-methoxyphenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.33 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.82-3.01, 3.47-3.63, 3.69-3.81,3.87, 3.87-4.00, 4.03-4.19, 4.23-4.34, 4.48-4.60, 5.14-5.27, 6.81-6.86,6.97-7.01, 7.06-7.16, 7.37-7.40, 8.23-8.27.

Example 19(34)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(3-methoxy-4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.31 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.82-3.01, 3.47-3.63, 3.69-3.81,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.48-4.60, 5.14-5.27, 6.90-6.97,7.00-7.04, 7.05-7.18, 7.32-7.41, 8.23-8.26.

Example 19(35)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3,4-dichlorophenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.37 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.50-3.63, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.44-4.55, 5.14-5.27, 6.92-6.98,7.12-7.22, 7.40-7.49, 8.21-8.26.

Example 19(36)6-amino-9-[(3R)-1-(2-butynoyl)-3-piperidinyl]-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.59 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.55-1.80, 1.84-2.11, 2.55-2.76, 3.07-3.21, 3.63-3.74,4.06-4.17, 4.38-4.59, 4.60-4.74, 7.04-7.23, 7.33-7.44, 8.20-8.24.

Example 19(37)6-amino-7-[4-(benzyloxy)phenyl]-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.22 (ethyl acetate);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.50-3.62, 3.68-3.79,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.40-4.50, 5.08-5.27, 7.07-7.16,7.32-7.46, 8.18-8.25.

Example 19(38)6-amino-7-[4-(1,3-benzodioxol-5-yloxy)phenyl]-9-[1-(2-butynoyl)-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.69 (ethyl acetate:methanol:aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.80-3.01, 3.47-3.62, 3.69-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.40-4.51, 5.12-5.25, 6.01, 6.53-6.59,6.60-6.63, 6.79-6.83, 7.04-7.13, 7.32-7.40, 8.20-8.23.

Example 19(39)6-amino-7-(4-anilinophenyl)-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.66 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.23-2.40, 2.80-3.01, 3.50-3.62, 3.68-3.80,3.87-4.00, 4.02-4.19, 4.23-4.34, 4.46-4.60, 5.12-5.25, 6.02, 7.00-7.18,7.22-7.40, 8.18-8.22.

Example 19(40)6-amino-7-(4-benzoylphenyl)-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.62 (ethyl acetate:methanol=8:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.41, 2.81-3.01, 3.52-3.63, 3.68-3.81,3.88-4.01, 4.03-4.20, 4.25-4.36, 4.55-4.65, 5.17-5.29, 7.49-7.68,7.81-7.86, 7.97-8.03, 8.25-8.29.

Example 19(41)6-amino-7-(4-benzylphenyl)-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.53 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.94-2.02, 2.23-2.39, 2.80-3.00, 3.50-3.61, 3.67-3.79,3.87-4.00, 4.02-4.18, 4.23-4.32, 4.44-4.52, 5.12-5.24, 7.18-7.42,8.19-8.23.

Example 19(42)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(phenoxymethyl)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.50 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.25-2.40, 2.82-3.01, 3.51-3.63, 3.69-3.80,3.88-4.01, 4.05-4.19, 4.25-4.34, 4.45-4.54, 5.11-5.27, 6.95-7.05,7.28-7.37, 7.43-7.51, 7.60-7.66, 8.22-8.27.

Example 19(43)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-{4-[(6-methyl-2-pyridinyl)oxy]phenyl}-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.46, 2.81-3.01, 3.50-3.62,3.68-3.80, 3.87-4.01, 4.03-4.19, 4.23-4.34, 4.52-4.62, 5.14-5.27,6.73-6.78, 6.93-6.98, 7.23-7.32, 7.40-7.46, 7.60-7.67, 8.21-8.25.

Example 19(44)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-fluoro-3-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.51 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.52-3.63, 3.68-3.80,3.87-4.01, 4.03-4.19, 4.23-4.34, 4.42-4.54, 5.14-5.27, 6.84-6.95,6.99-7.12, 7.34-7.41, 8.20-8.24.

Example 19(45)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-methoxy-3-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.51 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.23, 2.24-2.39, 2.81-3.01, 3.52-3.62,3.68-3.80, 3.85, 3.87-4.01, 4.03-4.19, 4.23-4.34, 4.42-4.54, 5.14-5.26,6.80-6.84, 6.85-6.93, 7.01-7.09, 7.31-7.39, 8.20-8.24.

Example 19(46)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-fluoro-3-methoxyphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.48 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.48-3.63, 3.68-3.80,3.87-4.01, 4.03-4.19, 4.23-4.34, 4.44-4.56, 5.14-5.26, 6.57-6.63,6.70-6.78, 7.03-7.14, 7.37-7.43, 8.20-8.24.

Example 19(47)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(3,4-dimethylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.53 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.48-3.62, 3.68-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.48-4.59, 5.14-5.25, 6.80-6.84, 6.89,7.04-7.17, 7.32-7.40, 8.20-8.24.

Example 19(48)6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-[4-(4-chloro-3-methylphenoxy)phenyl]-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.64 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.24-2.40, 2.81-3.01, 3.52-3.64, 3.70-3.80,3.87-4.00, 4.03-4.19, 4.23-4.34, 4.48-4.59, 5.14-5.25, 6.82-6.88, 6.98,7.06-7.15, 7.33-7.42, 8.20-8.24.

Example 19(49)6-amino-9-{(3R)-1-[(2E)-3-chloro-2-butenoyl]-3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

TLC: Rf 0.64 (ethyl acetate:methanol=19:1);

¹H-NMR (CDCl₃): δ 2.23-2.31, 2.32-2.40, 2.54-2.60, 2.86-2.97, 3.47-3.65,2.81-3.01, 3.52-3.64, 3.84-3.88, 3.93-4.01, 4.08-4.14, 4.20-4.25,4.40-4.54, 5.15-5.24, 6.20-6.31, 7.07-7.17, 7.18-7.22, 7.35-7.44,8.20-8.24.

Example 206-amino-9-{1-[(2E)-4-(1H-benzotriazol-1-yloxy)-2-butenoyl]-3-azetidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin-8-one

4-bromocrotonic acid (0.44 g),1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (0.86 g),HOBt (0.68 g), and triethylamine (1.3 mL) were added to adimethylformamide (22 mL) solution of the compound (1.0 g) prepared inExample 7, followed by stirring for 1 hour at room temperature. Waterwas added to the reaction mixture and extraction with ethyl acetate wasperformed. The organic layer was washed with a saturated aqueous sodiumchloride solution and was then dried over anhydrous sodium sulfate andthe solvent was distilled off. The residue was purified by silica gelcolumn chromatography to obtain the title compound (0.43 g) having thephysical property values indicated below.

TLC: Rf 0.56 (dichloromethane:methanol:28% aqueous ammonia=9:1:0.1);

¹H-NMR (CDCl₃): δ 4.43-4.63, 4.79-4.88, 4.99-5.05, 5.22-5.24, 5.35-5.45,6.26-6.34, 7.01-7.23, 7.36-7.44, 7.51-7.62, 8.22.

Example 216-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-{4-[hydroxy(phenyl)methyl]phenyl}-7,9-dihydro-8H-purin-8-one

A methanol (1 mL) solution of the compound (30 mg) prepared in Example19(40) was cooled to 0° C.; sodium borohydride (2.4 mg) was added; andstirring was carried out for 30 minutes. The reaction mixture wasdiluted with ethyl acetate and was then washed with water and asaturated aqueous sodium chloride solution. The organic layer was driedover sodium sulfate followed by concentration under reduced pressure,and the obtained residue was purified by silica gel columnchromatography (ethyl acetate→ethyl acetate:methanol=10:1) to obtain thetitle compound (28 mg) having the physical property values indicatedbelow.

TLC: Rf 0.58 (ethyl acetate:methanol=9:1);

¹H-NMR (CDCl₃): δ 1.94-2.03, 2.23-2.39, 2.57, 2.80-3.00, 3.50-3.61,3.67-3.79, 3.87-4.00, 4.02-4.18, 4.23-4.32, 4.45-4.54, 5.12-5.24, 5.91,7.27-7.43, 7.56-7.62, 8.20-8.23.

PHARMACOLOGICAL EXPERIMENTAL EXAMPLES Biological Example 1

Measurement (In Vitro Tests) of the Btk Inhibitory Activity and theSelectivity for Btk

The Btk enzyme inhibitory activity was measured, based on the protocolprovided by the manufacturer, using Btk (Invitrogen Corporation) and theZ′-LYTE™ Kinase Assay Kit-Tyr1 peptide (Invitrogen Corporation), whichcontained the following reagents: Tyr-1 peptide, Thy-1 phosphopeptide,5× kinase buffer, ATP, development reagent B, development buffer, andstop reagent.

5 μL/well of a solution of the test compound diluted with dimethylsulfoxide (DMSO), or DMSO, and 10 μL/well of the substrate/enzymemixture solution were dispensed to a 96-well assay plate and a reactionwas carried out for 20 minutes at 30° C. The substrate/enzyme mixturesolution was prepared by dilution with the kinase buffer(DL-dithiothreitol (DTT, 2.7 mM), 1.33× kinase buffer) to provide afinal concentration for the Tyr-1 peptide of 4 μM and a final Btkconcentration of 5 nM. 5 μL/well of the adenosine triphosphate (ATP,final concentration=36 μM) was then added and a reaction was carried outfor 1 hour at 30° C. After the completion of the reaction, 10 μL of adevelopment solution, provided by diluting the development reagent B to128× using the development buffer, was added and a reaction was carriedout for an additional 1 hour at 30° C. The enzymatic reaction was thenstopped by adding 10 μL of the stop solution. The fluorescence intensityat 445 nm and 520 nm in each well was measured using a Fusion UniversalMicroplate Analyzer (PerkinElmer Inc.) fluorescence plate reader. Thepercent phosphorylation was determined using the ratio of the emissionat 445 nm (coumarin emission) to the emission at 520 nm (fluoresceinemission) in accordance with the protocol provided with the kit.

The percent inhibition (%) by the test compound was calculated using thefollowing equation.

[E1]percent inhibition (%) of phosphorylation=1−{(A _(C) −A _(X))/(A _(C) −A_(B))}×100A_(X): % phosphorylation when the test compound has been addedA_(B): % phosphorylation in the absence of ATP addition (blank)A_(C): % phosphorylation when only DMSO has been added (control)

The 50% inhibition value (IC50 value) for the test compound wasdetermined from the inhibition curve based on the % inhibition at eachconcentration of the test compound.

The inhibitory activity for other kinases (for example, Lck, Fyn, andLynA (all from Invitrogen Corporation) was measured in the same manneras described above using the particular kinase in place of the Btk.

According to the results, the IC50 values for the compounds of thepresent invention were, for example, 0.004 μM for the compound ofExample 8, 0.014 μM for the compound of Example 11(3), 0.004 μM for thecompound of Example 8(14), 0.007 μM for the compound of Example 19(2),and 0.011 μM for the compound of Example 19(40).

In addition, the Btk-selective inhibitory activity of the compounds ofthe present invention for other kinases, and particularly for Lck, Fyn,and LynA, was calculated as the ratio of the IC50 values of theindividual kinases and is given in Table 1 below.

TABLE 1 Example Lck[IC50]/ Fyn[IC50]/ LynA[IC50]/ number Btk[IC50]Btk[IC50] Btk[IC50]  8 80 453 459  8(14) 220 2500 1767 11(3) 19 236 14319(2) 114 762 471

These results show that the compounds of the present invention not onlyhave a Btk inhibitory activity, but also have a Btk-selective inhibitoryactivity with respect to other kinases.

Biological Example 2

Measurement of B Cell Activation or T Cell Activation Using Human PBMC

A 10 mmol/L solution of the test compound in DMSO was dispensed into a96-well plate (Nunc) and a 5× dilution series was prepared using DMSO. A100× concentration test compound dilution solution was prepared by anadditional 10× dilution with RPMI1640 medium (contained 10% HI-FBS, 1%penicillin). Human peripheral blood mononuclear cells (PBMC) werediluted with the medium to provide a density of 2×10⁶ cells/mL. 396 μLof the cell suspension was added to a 96-well plate into which 4 μL ofthe 100× concentration test compound dilution solution or the solvent(10% DMSO) had already been introduced and incubation was performed for10 minutes at 37° C. and 5% CO₂. 10 μL of an anti-IgM antibody (SouthernBiotech)/IL-4 (R & D Systems) solution or an anti-CD3/CD28 antibody beadsuspension (Invitrogen Corporation) was added to a 96-well plate and 90μL of the cell suspension prepared as described above was also added(final concentrations: anti-IgM antibody=1 μg/mL, IL-4=3 ng/mL, andanti-CD3/CD28 antibody beads=2×10⁶ beads/mL). 10 μL of the medium wasadded to the unstimulated sample wells in place of these stimulatingsubstances, and standing at 37° C. and 5% CO₂ was again performed.Incubation was carried out for 16 hours in the case of the evaluation ofT cell activation and for 22 hours in the case of the evaluation of Bcell activation. 100 μL Cytofix Buffer (BD Biosciences) was added;holding for 15 minutes at 37° C. was performed; centrifugation for 10minutes at 1500 rpm was carried out; and the supernatant was removed.200 μL of Perm buffer II (BD Biosciences) at −20° C. was added; holdingfor 30 minutes on ice was performed; centrifugation for 10 minutes at1500 rpm was carried out; and the supernatant was removed. 0.5 mL Stainbuffer (BD Biosciences) was added and centrifugation was carried out for10 minutes at 1500 rpm. 100 μL of a mixed antibody solution was addedand incubation was performed for 30 minutes on ice in the dark. Theantibody was the 10× dilution with Stain Buffer of the 1:1:1 mixture ofPerCP-labeled anti-CD3 antibody (BD Biosciences), AF488-labeledanti-CD20 antibody (BD Biosciences), and PE-labeled anti-CD69 antibody(BD Biosciences). 0.4 mL Stain Buffer was added and the supernatant wasremoved. 0.3 mL Stain Buffer was added and the cell pellet was suspendedto prepare a sample for FACS measurement. The FACS analysis used a BDFACSCalibur (BD Biosciences) and CELLQuest Version 3.3 (BD Biosciences)data analysis software. The CD69-positive signal (average fluorescenceintensity) of the CD20-positive CD3-negative cells (B cells) orCD3-positive CD20-negative cells (T cells) was measured. Aftersubtracting the value for the unstimulated sample, the % inhibition withreference to the value for the stimulated control sample was determined.The % inhibition was graphically plotted using Prism (ver. 5.01J,GraphPad Software) and the IC50 value was determined.

According to the results, the IC50 values of compounds of the presentinvention for the CD69-positive signal for B cells was, for example,0.021 μM for the compound of Example 8, 0.032 μM for the compound ofExample 11(3), 0.023 μM for the compound of Example 8(14), and 0.061 μMfor the compound of Example 19(2). On the other hand, the IC50 values ofcompounds of the present invention for the CD69-positive signal for Tcells was >10 μM for all of the preceding compounds. Accordingly, thecompounds of the present invention were shown to have a selectiveinhibitory action on B cell activation.

Biological Example 3

Evaluation of the Stability in the Rat and Human Liver Microsome

(1) Preparation of the Test Compound Solution

A 0.25 mmol/L solution was prepared by diluting the test compound (10mmol/L DMSO solution, 5 μL) with an aqueous 50% acetonitrile solution(195 μL).

(2) Preparation of the 0-Minute Reaction Sample

245 μL of a 0.1 mol/L phosphate buffer (pH 7.4) containing 0.5 mg/mL ratand human liver microsomes (XenoTech) and NADPH-Co-factor (BDBiosciences) was added to a reactor that had been preheated to 37° C.;pre-incubation was performed for 5 minutes; and the previously indicatedtest compound solution (5 μL) was added and the reaction was started. 20μL was collected immediately after the start and the reaction wasstopped by the addition to 180 μL of acetonitrile that contained aninternal standard (warfarin). 20 μL of this solution was stirred with180 μL of an aqueous 50% acetonitrile solution on a filter plate forprotein depletion followed by suction filtration, and the filtrate wasused as the standard sample.

(3) Preparation of the 15-Minute Reaction Sample

The previously indicated reaction solution was incubated for 15 minutesat 37° C., and 20 μL was then added to 180 μL cold acetonitrile(contained the internal standard warfarin) to stop the reaction. 20 μLof this was stirred with 180 μL of an aqueous 50% acetonitrile solutionon a filter plate for protein depletion followed by suction filtration,and the filtrate was used as the standard sample.

(4) Evaluation Method and Results

The residual ratio (%) was calculated by injecting 1 μL of the samplesolution into an LC-MS/MS; dividing the peak area ratio for the reactionsample (peak area for the test compound/peak area for the internalstandard) by the peak area ratio for the standard sample; andmultiplying the resulting value by 100.

The following were used as test compounds: compounds of the presentinvention and1-{(3R)-3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-piperidinyl}-2-propen-1-one(comparative compound A) and1-{3-[4-amino-3-(4-phenoxyphenyl)-1H-pyrazolo[3,4-d]pyrimidin-1-yl]-1-pyrrolidinyl}-2-propyn-1-one(comparative compound B), which are exemplary compounds described inPatent Document 1 that have a pyrazolopyrimidine skeleton. The residualratio (%) of the test compound in the rat and human liver microsome wasas shown in Table 2 below.

TABLE 2 Residual ratio (%) Residual ratio (%) in the rat liver in thehuman liver Compound microsome microsome Comparative compound A 0 0Comparative compound B 50.6 55.9 Example 8 100 87.0 Example 19(2) 86.4100 Example 19(9) 100 97.9 Example 11(24) 88.5 100

The results show that the compounds of the present invention are muchmore stable in rat and human liver microsomes than the comparativecompounds.

Biological Example 4

Measurement of the Inhibitory Activity on Drug-Metabolizing Enzymes(Inhibitory Action on Human CYP2C8 and Human CYP2C19)

[Experimental Method]

The reaction was carried out in a 96-well plate. The positive controlsubstances (CYP2C8: ketoconazole, CYP2C19: tranylcypromine) wereadjusted (CYP2C8: 0.6 and 6 mmol/L, CYP2C19: 0.9 and 9 mmol/L) with DMSOto concentrations that were 300 times the final concentration, andsolutions were prepared (CYP2C8: 8 and 80 μmol/L, CYP2C19:12 and 120μmol/L) by 75× dilution with purified water that contained 2.7%acetonitrile. The test compounds were adjusted to 0.3 and 3 mol/L withDMSO and were adjusted to 4 and 40 μmol/L by 75× dilution with purifiedwater that contained 2.7% acetonitrile. A reaction mixture was thenprepared (the numerical values are final concentrations) by the additionof potassium phosphate buffer (pH 7.4), magnesium chloride (5 mol/L),substrate (CYP2C8: dibenzylfluorescein 1 μmol/L, CYP2C19:3-cyano-7-ethoxycoumarin 25 μmol/L), and E. coli-expressed livermicrosome CYP2C8 (Cypex, 10 pmol/L) and CYP2C19 (Cypex, 3 pmol/L). 100μL of this reaction mixture and 50 μL of the test compound prepared asdescribed above and the positive control solution prepared as describedabove were dispensed into each well and pre-incubation was carried outat 10 minutes for 37° C. The reaction was started by the addition of 50μL NADPH solution (final concentration=1 mmol/L) and incubation wascarried out for 30 minutes at 37° C. The fluorescence intensity wasmeasured (CYP2C8: excitation wavelength=485 nm, fluorescencewavelength=538 nm; CYP2C19: excitation wavelength=409 nm, fluorescencewavelength=460 nm) immediately after the addition of the NADPH and afterincubation for 30 minutes. The % inhibition was taken to be the %decline (% inhibition) in the fluorescence intensity in comparison to acontrol in which DMSO was added in place of the test compound solutionand the reaction was carried out and was calculated using the followingformula.inhibition (%)=100−{(fluorescence intensity after the reaction of thetest compound−fluorescence intensity prior to the reaction of the testcompound)/(fluorescence intensity after the controlreaction−fluorescence intensity prior to the control reaction)×100}

The IC50 value was taken to be <1 μM when the % inhibition at 1 μmol/Lwas at least 50%; was taken to be >10 μmol/L when the % inhibition at 10μmol/L was not more than 50%; and in between the preceding (not morethan 50% at 1 μmol/L and at least 50% at 10 μmol/L) was calculated usingthe following formulaIC50=(50−b)/awherein a and b are the slope and intercept of the linear regressionline y=ax+b that passes through the following two points: theconcentration of 1 μmol/L, % inhibition and the concentration of 10μmol/L, % inhibition.

The IC50 values of the comparative compounds and compounds of thepresent invention were measured using the measurement method describedabove.

The results were as follows: for comparative compound A and comparativecompound B, the IC50 values for CYP2C8 were 4.7 μM and 6.9 μM,respectively, and the IC50 values for CYP2C19 were 5.6 μM and 8.1 μM,respectively. On the other hand, for the compounds of the presentinvention, for example, the compounds of Example 8, Example 11(3),Example 8(14), and Example 19(2), all had IC50 values for CYP2C8 andCYP2C19 of >10 μM. Accordingly, the compounds of the present inventionwere shown to have a lower CYP inhibitory action than the comparativecompounds.

Biological Example 5

Measurement of the Cytotoxicity and Ability to Reduce the MitochondrialMembrane Potential in Cultured Human Hepatoma Cells

Tissue that maintains an aerobic equilibrium, e.g., the kidneys andheart, and tissue that is exposed to high drug concentrations andcarries out drug metabolism, e.g., the liver, are known to be sensitiveto mitochondrial dysfunction (Drug Discovery Today, 12 (17-18), 777-785,2007). The reduction or extinction of the mitochondrial membranepotential by a drug is caused by the direct inhibition of the electrontransport system, decoupling of electron transport from ATP synthase, orthe opening of a mitochondrial membrane permeability transition pore. Asa consequence, measurement of the mitochondrial membrane potential ofliver cells can provide a parameter for hepatotoxicity.

Human liver cells were seeded into a collagen-coated 96-well plate at acell density of 30,000 cells/well and incubation was carried outovernight in an incubator at 37° C. in 5% CO₂-95% air. The culturedcells were stained for 1 hour with5,5′,6,6′-tetrahydro-1,1′,3,3′-tetramethyl-benzamidazolocarbocyanineiodide (JC-1) followed by treatment with the test compound. The testcompound was dissolved in DMSO and then diluted with the liquid culturemedium Hepatocyte Culture Medium (HCM) and added to the cells. The testcompound treatment concentrations were 0, 6.25, 12.5, 25, 50, 100, 200,and 400 μmol/L. After exposure to the test compound for 24 hours,measurement was performed with a SpectraMax plate reader (MolecularDevices, LLC) at an excitation wavelength of 485 nm and a fluorescencewavelength of 538 nm and an excitation wavelength of 544 nm and afluorescence wavelength of 590 nm. The membrane potential was determinedby the ratio between the 544 nm/590 nm measurement value and the 485nm/538 nm measurement value. After this, the ATP concentration in thecells was measured using a Celltiter Glo luminescent assay kit (PromegaCorporation) in order to evaluate the cell toxicity of the testcompound. The cells were lysed by the assay buffer provided with themeasurement kit and the concentration of the ATP released from the cellswas measured using the luciferin-luciferase enzyme activity as theindex. The emission was measured using a SpectraMax plate reader. Theability of the test compound to lower the mitochondrial membranepotential and the cytotoxicity of the test compound were represented bythe concentration (IC50 value) that caused a 50% decline in themitochondrial membrane potential and ATP concentration, respectively.The ability to lower the mitochondrial membrane potential of the testcompounds and the cytotoxicity of the test compounds are given in Table3 below.

TABLE 3 Ability to lower the mitochondrial Toxicity for human membranepotential liver cells (IC50 Compound (IC50 (μM)) (μM)) Comparativecompound A 39 78 Comparative compound B <6.25 <6.25 Example 8 324 290Example 19(2) 200 135 Example 19(9) 181 97 Example 11(24) 347 213

The results showed that both of the IC50 values were lower for all ofthe compounds of the present invention than for the comparativecompounds.

FORMULATION EXAMPLES Formulation Example 1

The components indicated below were mixed by a standard method, filteredacross a dedusting filter, filled into 5 mL ampoules, and thermallysterilized with an autoclave to obtain 10,000 ampoules that contained 20mg active component per ampoule.

6-amino-9-{(3R)-1-[(2E)-4-(dimethylamino)-2-butenoyl]- 200 g3-pyrrolidinyl}-7-(4-phenoxyphenyl)-7,9-dihydro-8H-purin- 8-one mannitol 20 g distilled water  50 L

Formulation Example 2

The components indicated below were mixed by a standard method and thentabletted to obtain 10,000 tablets that contained 10 mg of the activecomponent in each tablet.

6-amino-9-[(3R)-1-(2-butynoyl)-3-pyrrolidinyl]-7-(4- 100 gphenoxyphenyl)-7,9-dihydro-8H-purin-8-one calcium carboxymethylcellulose (disintegrant)  20 g magnesium stearate (lubricant)  10 gmicrocrystalline cellulose 870 g

INDUSTRIAL APPLICABILITY

The compounds of the present invention are compounds that, in additionto having a Btk-selective inhibitory activity, exhibit an excellentmetabolic stability and can avoid hepatotoxicity or the like, and as aconsequence are useful as very safe therapeutic agents for diseases inwhich B cells or mast cells participate.

The invention claimed is:
 1. A method of preparing a compoundrepresented by general formula (I-D):

wherein L represents (1) —O—, (2) —S—, (3) —SO—, (4) —SO₂— (5) —NH—, (6)—C(O)—, (7) —CH₂—O—, (8) —O—CH₂—, (9) —CH₂—, or (10) —CH(OH)—; R¹represents (1) a halogen atom, (2) a C₁₋₄ alkyl group, (3) a C₁₋₄ alkoxygroup, (4) a C₁₋₄ haloalkyl group, or (5) a C₁₋₄ haloalkoxy group; ring1represents a 4- to 7-membered cyclic group, which may be substituted byfrom one to five substituents each independently selected from the groupconsisting of (1) halogen atoms, (2) C₁₋₄ alkyl groups, (3) C₁₋₄ alkoxygroups, (4) nitrile, (5) C₁₋₄ haloalkyl groups, and (6) C₁₋₄ haloalkoxygroups, wherein when two or more substituents are present on ring1,these substituents may form a 4- to 7-membered cyclic group togetherwith the atoms in ring1 to which these substituents are bonded;ring2^(d) represents a 4- to 7-membered nitrogenous saturatedheterocycle substituted by at least one —K^(b)—R^(2d); K^(b) represents—C(O)—; R^(2d) represents a C₁₋₄ alkylene, C₂₋₄ alkenylene, or C₂₋₄alkynylene group which may be substituted by CONR⁵R⁶, CO₂R⁷, or OR⁸; R⁵and R⁶ each independently represent (1) a hydrogen atom, (2) a C₁₋₄alkyl group, or (3) a phenyl group; R⁷ represents (1) a hydrogen atom or(2) a C₁₋₄ alkyl group; R⁸ represents (1) a hydrogen atom, (2) a C₁₋₄alkyl group, (3) a phenyl group, or (4) a benzotriazolyl group; nrepresents an integer from 0 to 4; m represents an integer from 0 to 2;and when n is two or more, the R¹'s may be the same as each other or maydiffer from one another, or a salt thereof; wherein the methodcomprises: reacting a compound represented by general formula (I-A):

wherein ring2^(a) represents an unsubstituted 4- to 7-memberednitrogenous saturated heterocycle, while the other symbols have the samedefinitions as above, with a compound represented by formula (I-D-1)R^(2d)—CO₂H  (I-D-1) wherein R^(2d) is defined as above, using acondensing agent.
 2. The method of preparing a compound according toclaim 1, wherein ring1 is a benzene, cyclohexane, or pyridine ring, eachof which may be substituted by from one to five substituents eachindependently selected from the group consisting of (1) halogen atoms,(2) C₁₋₄ alkyl groups, (3) C₁₋₄ alkoxy groups, (4) nitrile, and (5) CF₃.3. The method of preparing a compound according to claim 1, wherein the4- to 7-membered nitrogenous saturated heterocycle is an azetidine,pyrrolidine, or piperidine ring.